Lakhtin V., Lakhtin M., Aleshkin V. G.N. Gabrichevsky Research Institute for Epidemiology and Microbiology, Russia INTERACTION OF WITH CARBOHYDRATE-SENSITIVE REAGENTS Summary. The data on glycobiological aspects of 63 esterases (interacting with carbohydrate- sensitive proteins and other reagents) from 360 sources are systemized and presented as a table. Key words: esterases, glycosidases, lectins, glycoconjugate recognition, biotechnology, microbiology, medicine. Isolation of esterases using lectins Lectin chromatography step, as the most effective one, provided significant purification (30- 1,000 fold [f]) in the following cases of esterases: 9-O-acetylated sialate-specific from the rat liver microsomes (31 f) [451]; pseudocholinesterase of fish (28 f) [424]; human milk (111 f) [377], rat liver lysosomal A1 (165-1,000 f) [440, 441]; human plasma (85 f) [447]; bovine mammary gland microsomal alkaline (30 f) [471]; from the sunflower seeds or mammalian tissues (28-40 f) [618, 556, 601]; bovine liver cytosolic 5'- (33 f) [630]; soybean nucleotide-specific phosphatase (123 f) [669]; human tissue acid sphingomyelinase (54-108 f) [708, 709, 705]; human urine arylsulphatase A (119 f) [741]; mammalian liver arylsulphatase B (263-465 f) [750]; human placental N-acetylgalactosamine-6- sulphate sulphatase (64 f) [733]; SP from the spinach leaves (37 f) [810]; the differentiation-specific nuclease from the plant mesophyll cells (72 f) [808]; and fish 5'-inosinate- monophosphate (157 f) [593]. It should be noted that in some cases the sequential chromatography (the use of coupled column sorbents including immobilized lectin and other non-lectin type sorbent or sorbents) was especially effective and suitable for isolation of the following esterases: human milk lipase, 111 f (filtration through ConA-sorbent followed by binding to cholate-sorbent, ConA-Cholate+) [377]; rat tissue lysosomal (combination of hydroxyapatite and DEAE-cellulose with ConA- sorbent (HA—DEAEC—ConA) [398], or Orange and Red dye agaroses with ConA- and phenyl- agarose [442], or hydroxyapatite with ConA-sorbent (HA—ConA), 254 f [438]); mammalian tissue arylphosphatases, 104-163 f (immobilized ConA and Blue-dye-agarose) [751, 761]; mammalian tissue iduronate- or glucuronate-2-phosphatase (immobilized ConA and Blue-dye-agarose, 119-853 f [756, 762], or immobilized ConA, Blue agarose and Green agarose [757, 1735, 1734]). A lot of examples for effective separation of esterase multiple forms using lectin sorbents can also be found in the table given below. Some esterases reveal relatively high hydrophobic properties so that ethylene glycol is needed for successful affine chromatography of using lectins [615, 622]. In some cases significant activation resulted in high purification levels of esterases using lectin sorbent alone or coupled with other type of sorbent: rat liver lysosomal phospholipase A1 (56% of activation, 165 f) [440]; rat kidney cortex lysosomal phospholipase A1 (130% of activation, 254 f) [438]; human liver N-acetylglucosmine 6-sulphate sulphatase (110% of activation, 163 f) [761]. Slight activation was observed during lectin chromatography of mammalian carboxyesterase [365], acid phosphatase [606], 5'-nucleotidase [657], or acid sphingomyelinase [709]. It should be noted that activation of esterases may depend on isoform spectrum and the concrete isozyme selected for purification using lectin chromatography [365]. Carbohydrate moiety of esterases interacting with carbohydrate-binding proteins The contents of sugars for esterases are significantly varied as it can be seen from the table: from less than 1% (w/w) for wheat germ acid phosphatase [579] up to 50-55% for lower plant

[394] and acid [536, 542-543, 547], or higher plant acid phosphatases [548, 620]. In majority of cases, esterases have varied N-linked glycans (see the table). carbohydrate moiety in some cases can be associated with polysaccharides: for example, O-linked mannan of yeast acid phosphatase [539], or chitin associated with minor form of insect [462]. Other types of carbohydrate moiety can be O-linked clusters (mucin-like) of residues of GlcNAc as in human nuclear protein tyrosine phosphatase [681], or clusters of sialic acid residues sensitive for sialic acid-binding proteins [415]. The use of such inhibitors of intracellular processing glycosylation of proteins as tunicamycin [361, 449, 443, 546, 444, 459] or swainsonine [522] allows obtaining deglycosylated esterase forms and, therefore, evaluation of the size of enzyme carbohydrate moiety. For example, rat heart monomeric extracellular phospholipase contains 12,5% sugars (7 kDa within 56 kDa) compared to intracellular form from tunicamycin-treated cells [449]; chicken or bovine adipocyte lipoprotein lipase contains 12,6 or 15,8% sugars, respectively (that means the presence of three Asn-linked glycans in the monomeric enzyme) [443, 444]. Endoglycosidase-treatments can be also used for calculation of sugar contents. Thus, rat liver microsomal carboxylesterase precursor includes 3,2% sugars sensitive for EndoH-splitting (Asn- glycans of oligomannoside type of EndoH+-enzyme form) [361, 362]; fungal lipase includes 8,1% sugars sensitive for EndoF (EndoF+-enzyme form) [369]; yeast lysophospholipase EndoH+-forms 220 and 145 kDa include 69 or 53% sugars [392]; shrimp or human liver alkaline phosphatase EndoF+-forms 50 and 75 kDa contain 22 or 33% of Asn-glycans [13, 465]; yeast acid phosphatase (EndoF+) 140 kDa includes 60% Asn-glycans of oligomannoside type [545]; leishmanial promastigote lysosomal acid phosphatase (EndoF+) 70-72 kDa includes about 30% Asn-glycans [586], and another leishmanial 3'-nucleotidase 43 kDa contains about 3% N-glycanase-sensitive glycans [671]; human protein tyrosine phosphatase 114 kDa (expressed in insect cells SF-9) includes 14% Asn-glycans of complex type [679]; murine plasmacytoma cell I (115 kDa) includes 10% N-glycanase-sensitive Asn-glycans [695]; and fungal (having subunits of three types) includes glycosylated polypeptides 34 and 30 kDa (21 or 8% Asn-glycans of complex type, respectively) [796]. Combinative and/or sequential glycosiadases-treatments allow deeper knowledge on glycoprotein esterases. Other endoglycosydases can be useful in study of esterases: EndoD in case of bovine 5'- nucleotidase [628], or endo-beta-galactosidase in the case of human ribonuclease [803]. In addition, the treatments of esterases with alpha-mannosidases [364, 414, 628, 780, 729], glucosidases [571, 653], beta-galactosidase [571, 628], alpha-fucosidase [571, 628], beta-N-acetylglucosaminidase [729, 628], lysozyme [729], or cellulolysin including cellulase and hemicellulase [565] were used. A lot of examples for the use of sialidase-treatments of esterases (especially for mammalian enzymes) can be found in table. As a result of such treatment, human tissue alkaline phosphatasees or rat liver 5'-nucleotidase isoforms became more cationic [497, 664] and included more restricted spectrum of forms [664]. More over, simple calculation of content of sialic acid residues in esterase treated with sialidase can be made [664]. Glycosidase-treatments can result in alteration of esterase binding to a set of lectins. For example, EndoH-treated rat brain reveals low affinity to immobilized ligatin [422]; bovine ribonuclease-B treated with endoglycosidases loose ability to interact with ConA or WGA [787, 788]; fungal treated with EndoH does not bind to ConA-sorbent [806]. Sialidase-treatment results in significant decreasing of the bovine acetylcholinesterase binding to immobilized WGA [414] and does not allow further precipitation of the human erythrocyte with this lectin [415]. However, EndoH-resistant (EndoH-) sialidase-treated form of guinea pig adipocyte lipoprotein lipase reveals increased binding ability with respect of RCA-I-

2 sorbent [446]. Interestingly, sialidase-treatment of human tissue alkaline phosphatase decreases enzyme precipitation with WGA, but significantly increases precipitation with SBA [210]. Simultaneous increasing affinity to PNA and decreasing affinity to WGA were observed for sialidase-treated frog liver acid phosphatase forms [596]. Thus contribution of sialic acid residues of O-linked glycans of mucin type into interaction between esterase molecyles and lectins takes place [210, 495, 489, 596]. Sequential glycosidase-treatments of esterases allow to study internal sugar residues in glycans, as it can be seen in the cases of bovine 5'-nucleotidase [628], or human ribonuclease and arylsulphatase [739, 803]. Additional lectin analysis of all esterase forms modified during sequentional splitting of sugars allows both control of each glycosidase type effectivity and preliminary suggestion of the whole glycan structures [628]. A set of lectins (mainly lectin sorbents) can be very effective for separation and classification (lectin typing) of esterase forms, as it was demonstrated for example for rat tissue cholinesterase [423, 426]; mammalian alkaline phosphatases [210, 479, 487, 489, 493, 497, 513, 514, 518, 522]; frog or rat liver acid phosphatases [596, 616]; human liver acid sphingomyelinase acid or neutral forms [707]; human -I forms [767, 768]; and human urine ribonuclease-U forms [802]. The direct chemical analysis of carbohydrate moiety of esterases is usually in accordance with lectin analysis or results: for example, in cases of porcine pancreatic lipase [379, 380], rat liver microsomal acetylesterase [368], silkworm alkaline phosphatase [461], mammalian alkaline phosphatase (see table), or human [802, 803]. The patterns of separation of glycopeptide or glycan mixtures from esterases using a set of lectin sorbents supported expectable behaviour of intact esterases during lectin chromatography: for example, in cases of such mammalian extensively studed enzymes as , , alkaline phosphatases, and . The use of endo- and exoglycosidases makes lectin analysis more reliable, and establishment of chemical structure of isolated esterase glycans using modern chemical or physical methods should use for the final conclusions about the detailed carbohydrate moiety organization. The final elucidation of glycan structures is especially important for unusual, unique and/or modified with groups other than sugar residues. Sometimes, phosphorylated or sulphated Asn- glycans can be found in esterase molecules [79, 882]. Sulphated Asn-glycans keep their sensitivity for splitting by EndoH or EndoF [422, 443], or their ability to interact with ConA [443]. Phosphorylated Asn-glycans are essential for interaction between rat brain alkaline phosphatase and endogenic lectin (ligatin) [422], or for targetting of enzymes into mammalian lysosomes through recognition of mannose-6-phosphate-residues of enzymes by receptor lectin [1848, 1849]. Glycolipids can be included in esterases upon their solubilization from cell membranes. In the latter cases, the treatment of esterase with can result in increasing enzyme affinity to ConA [627]. It was found that insect possesses Fuc-alpha-1,3-residues in the core position of Asn-glycans similarly to plant but not vertebrate glycoproteins [389]. Non-enzymatically glycosylation of calf intestinal alkaline phosphatase or bovine ribonuclease may result in additional enzyme sites recognized by carbohydrate-binding proteins [7, 780]. In some cases carbohydrate moiety of esterases is important for enzyme activity [7, 544, 653, 729]. Thus, non-enzymatic glucosylation of the bovine alkaline phosphatase results in partial inactivation [7]. Exoglycosidase-treatments can also serve as inactivators of esterases, as it was shown for human placental sulphatases [729] and rat brain 5'-nucleotidase [653]. Interestingly, elimination of EndoH-sensitive glycans resulted in activation of the yeast lysophospholipase [920]. It seems, carbohydrate moiety can be regulated by expressing of esterase gene in the host

eucaryotic cells, as in the case of human phosphotyrosyl- expressed in insect cells [679], or ribonuclease gene products expressed in tobacco cells [797]. General comments for the following table on esterases Left column Enzymes are listed according to Enzyme Commission (EC) numbers ([1791], completed). Sources of enzyme are given in the following order: viruses, bacteria, lower plants (yeasts, fungi), higher plants, invertebrates, vertebrates (including mammals). Within each group pointed above sources are given according to alphabetical order of latin genus and species (alphabetical order of common names is possible). Molecular or SU masses are pointed according to methods of their determination (in brackets). Sequential procedures or conversion processes are pointed. The most effective step of enzyme purification is marked as **, and steps of HPLC, FPLC, HPLAC are marked as *. Calculated carbohydrate moiety is also given if possible. Right column Lectin+ means result of interaction between lectin and enzyme or influence toward enzyme activity (complex, precipitate, sorption of enzyme to lectin sorbent, retardation of enzyme during lectin chromatography or LIE, inhibition, activation, immobilization and stabilization of enzyme). Relative affinity of enzyme to lectin can be also marked (2+,3+, coefficients) in the case of a panel of lectin tested. EndoH+, EndoF+, PNGase+ or Glycosidase+ means specific splitting of GP. Sensitivity of carbohydrate moiety for stain is also shown (PAS+, others).

Table Interaction of esterases and their protein modulators with carbohydrate-sensitive reagents

Enzyme, sources, procedures, Carbohydrate-sensitive Ref. carbohydr. moiety, comments reagents, comments

3.1.1.1. Carbohylesterase

Lower plant Aspergillus niger (fungal extract) Serine carboxylesterase 120kDa, SU 60kDa, pI 4,5 CMC**--Octyl/Phenyl-Sepharose— SephadexG150 + Carbohydr.: 4% (Phenol- H2S04) [357]

Mammals Dog and human pancreas Monomeric carboxylesterase forms Potential Reagents+ 87 and 93kDa (dog), and 100kDa (human) Ab(anti human enzyme)-Affigel10 Carbohydr.: 4,8% (93kDa), 5,8% (100kDa) [358]

Porcine liver microsomes Carboxylesterase forms 58,2+59,7+ Potential Reagents+ 61,4kDa, pI 4,8-5,0 GlcN (M/M/): 1,8 (58,2kDa), 1,7 (61,4kDa) [359]

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Rabbit liver Carboxylesterase 183kDa,SU 63kDa, pI 5,3-6,0 DEAEC—SephadexG150 + Carbohydr. in 63kDa (Thymol-H2S04) [360]

Rat hepatocytes in culture Carboxylesterase mature and precursor forms 61-62kDa, and deglycosylated form from tunicamycin-treated cells Ab-precipitation—ConA-Sepharose ConA+ Asn-glycans of oligoMan-type (2kDa EndoH+ (62—60kDa) in 62kDa) [361]

Monomeric carboxylesterase 60kDa, pI 6,1; precursor 59kDa; and deglyco- sylated form 58kDa Prepar.IEF—DEAE-Trisacryl—Superose12*-- ConA-Sepharose ConA+ (desorption of 60 kDa with 0,2M MM; separation from 59kDa) Asn-glycans of oligoMan-type (2kDa EndoH+ (60—58kDa), in 60kDa) EndoH- (59kDa) [362]

Rat intestine Cefuroxime axetil carboxylesterase 51kDa, SU 31,5+26,8kDa, pI 4,5-6,3 DEAEC**—UltrogelAcA34—HA—ConA- ConA+ (desorption with Sepharose 0,3M MM using sugar gradient; 48%, 1,2-f) [363] Rat liver cell microsomal lumens (endoplasmic retuculum cisternal space) Soluble carboxylesterase E1 175kDa, SU 59kDa, pI 5,7 ConA-Sepharose—SephadexG200— ConA+ (desorption with DEAEC—HA 0,2M MM) Carbohydr. in 59kDa (M/M): 8 hexoses, 2 GlcN(Ac); Asn-glycans: oligoMan type (2kDa) EndoH+, alpha-mannosidase+ [364]

Monomeric carboxylesterase 57-60kDa forms pI 5,0+5,5+6,4, and trimeric form pI 6,1 IEF—HA—DEAEC—ConA-Sepharose6B ConA+ (pI 5,0, 6,1, or 6,4; desorption with 0,5M MM; 104%, 2,9-f, for pI 5,0) Affinity to 125I-ConA ConA+ (for 3 forms) [365]

Monomeric carboxylesterase 62kDa, pI 6,4 Prapar.IEF—DEAE-Trisacryl— Superose12*--ConA-Sepharose ConA+ (desorption with 0,2M MM) [361] 3.1.1.2. Arylesterase

Horse serum

Esterase SDS-PAGE—Blot—Lectin-biotin ConA+, PNA+ [366]

Human serum Paraoxonase 43kDa (37kDa for Potential Reagents+ deglycosylated form) Carbohydr.: 15% [367]

Rat liver microsomes Monomeric arylesterase (A-esterase) 61kDa ConA-Sepharose**—SephacrylS300— ConA+ (80%, 3,0-f) DEAE-Sepharose—HA—IEF Carbohydr.(M/M): 7.4 Man, 2.5 GlcN; 11 variants of Asn-glycans of oligoMan type [368]

3.1.1.3. Triacylglycerol lipase

Lower plants Galactomyces geotrichum CBS 772.71 (fungal CF) Monomeric lipase 62kDa, pI 5,2 Asn-glycans: 5kDa EndoF+ (62—57kDa) [369]

Geotrichum candidum (fungal CF) Lipase forms 64 and 66kDa + Carbohydr.: 6,.5-6,6% (Phenol-H2S04) [370]

Mucor mihei (fungal CF) Monomeric tributyrolglycerol hydrolase forms A (32kDa), and B (deglycosylated A) DEAEC**--ConA-Sepharose ConA+ (70%, 2-f, for A) [371] Oospora lactis (micromycete, CF) Purified lipase, 8% carbohydr.: 8% (75-80% Man hexoses) [372]

Rhizopus arrhizus (mold, CF) Lipase-I 42,7kDa (SephadexG100) associated with Gp (8,5kDa) Carbohydr.: 13-14 Man, 2 hexosamines [373]

Mammals Human foreskin fibroblast CF Monomeric lysosomal acid lipase/ cholesterol esterase 47-49kDa Decyl-agarose**--UltrogelAcA44—HA— Phenyl-boronate-gel—UltrogelAcA44— UltrogelAcA44; ConA-agarose ConA+ (desorption with MM) Asn-glycans: oligoMan type (4-6kDa) EndoH+ [374]

Human liver lysosomes Acid lipase 125kDa, SU 29kDa ConA-Sepharose**--CMC—SephadexG150 ConA+ (desorption with 0,18M MM; 48%, 11-f) [375]

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Monomeric acid lipase/cholesterol esterase forms 2 (58,5-69kDa), and 1 (29kDa) ConA-Sepharose**--CMC—MonoS* ConA+(binding of 90%; desorption with 0,37M MM; 71%, 4,7-f) [376] Human milk Bile-salt-activated monomeric lipase 125kDa ConA-Sepharose4B—Cholate- ConA-(separation from Sepharose bound GP),ConA-Cholate+ (57%,111-f) [377] Carbohydr.: 9.2% [358]

Human plasma Lipase 75kDa (SephadexG150) or 64kDa PAS+ (PAGE), pI 4,1,associated with phospholipase A1 Heparin-Sepharose**--ConA-Sepharose— ConA+ (desorption with Column-IEF 1M MG; 81%, 3,9-f) [378]

Porcine pancrease Lipase forms A and B SephadexG100—DEAEC—CM-Sephadex; SephadexG100—DEAEC—ConA-agarose ConA+ (both forms; binding of 99%; desorption of 36% with 0,5M MM) Carbohydr.(M/M): 4 Man, 3 Glc(NAc); [379] Asn-glycans of oligoMan-type and fucosylated biantennary of complex type [380]

Rabbit gastric tissue Tributyrin-specific lipase 49kDa (SDS- PAGE), forms pI 5,7-7,0 Sephadex- Carbohydr.: 17-18% [381]

Rabbit liver lysosomes Monomeric acid lipase 42kDa solubilized with digitonin BiogelA1.5m—DEAE-Biogel—Phenyl- Sepharose**--Prepar.PAGE—Affigel-Blue (or ConA-Sepharose) ConA+(desorption with 0,75M MM) [382] Rat adrenal gland and liver Microsomal lipase precursor 51kDa and extrahepatic mature form 53kDa Asn-glycans of oligoMan-type (in 51 and EndoH+ (51kDa), 53 kDa), and of complex type (in 53kDa) EndoF+ (53 and 51kDa) [383] Rat liver lysosomes Monomeric trioleinate esterase 68kDa solubilized with TX100 ConA-Sepharose**--SephadexG150 ConA+ (desorption with 0,5M MM; 61%, 15-f) [384]

3.1.1.4. Phospholipase A2 Insect Apis mellifera (honeybee venom) Enzyme forms 16, 18, 20kDa (SDS-PAGE) BiogelP60—ConA-Sepharose4B— ConA- (12%), ConA+ BiogelP10 (desorption of 33% with 15mM, and residual 55% with 0,5M MM) [385]

ConA-Sepharose4B—Prepar.PAGE ConA+ (20kDa), ConA- (16, 18kDa),ConA- (38%), ConA+ (desorption with 0,5M MM) [386,387] Mixture of Gp of 18, 20kDa ConA-Sepharose ConA+ (separation of Gp) 4 Asn-glycans with terminal Fuc and PNGaseF+ ([20+18]—16kDa), GlcNAc; GalNAc in 20kDa EndoF+ ([20+18]—16kDa), EndoH+ (20kDa),EndoD+ (18kDa), EndoD- (20kDa), alpha-Mannosidase+ (18—16 kDa), alpha-L-Fucosidase+ [385,386,388,389] Mammal Bovine adrenal medulla PhospholipaseA2 30,6kDa, pI 7,8+8,1+8,4 DEAEC**—ConA-Sepharose4B— ConA+(complete binding; SephacrylS200 desorption with 0,5M MM; 56%, 27-f) [390] 3.1.1.5. Lysophospholipase

Lower plants Penicillium notatum (fungus) Purified phospholipaseB 95kDa (SDS-PAGE), 603aar (64779 Da) Carbohydr.:approx. 24,5kDa; 16 potential Potential Reagents+ Asn-glycosylation sites; Man9GlcNAc2-Asn [391]

Saccharomyces cerevisiae H1022 (yeast CF, plasma Mb)

Extracellular phospholipaseB 200-280kDa, pI 4-6, 41 mkM sugars/mg-protein SephacrylS300—ConA-Sepharose (or ConA+ Chromatofoc) [392]

Monomeric plasma Mb-bound phospholipase B forms 220 and 145kDa Carbohydr.: 22 or 10 mkM hexoses/mg EndoH+ (220+145—67kDa), protein, for 220 or 145kDa, (EndoH+EndoF)+ (220— respectively 52kDa) [393]

Torulaspora delbrueckii (Saccharomyces rosei, Yeast CF) PhospholipaseB 170-200kDa (diffuse band,

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SDS-PAGE) Carbohydr.: approx.50%, Asn-glycans of oligoMan type (75% of EndoH+ ([170-200]—72kDa) total sugars) [394]

Higher plants Hordeum vulgare (barley germinating seedlings) Monomeric lysophospholipase 40kDa, CMC—Octyl-agarose**—ConA-agarose— ConA+ (desorption with Phenyl-agarose—BiogelP100—BiogelP100 1M NaCl; 44%, 2,9-f) Carbohydr.: 10-12% hexoses [395]

Solanum tuberosum cv. Russet Burbank (potato tubers and leaves) Phospholipase (C6-NBD-PC hydrolase) SephadexG50—DEAEC—ConA-Sepharose ConA- (72%; leaf enzyme), ConA+ (desorption of 23% with 20mM MG, and residual 5% with 0,3M MG, leaf enzyme; desorp- tion of 45 and 55% with 20mM and 0,3M MG, respectively, for tuber enzyme) [396] Mammals Rat intestinal brush-border Mb PhospholipaseB 120kDa associated with phospholipaseA2 ConA-Sepharose ConA+ Aminosugars: 70% GlcNAc, 30% GalNAc [397]

Rat liver lysosomes Soluble lysophospholipase associated with phospholipaseA1 HA—DEAEC—ConA-Sepharose— ConA+ (complete binding; Chromatofoc desorption with 0,5M MM) [398] Rat platelets activated with thrombin Monomeric extracellular lysophospholipase 32kDa (SDS-PAGE) or 47kDa (TSKgelG2000SW)* Heparin-Sepharose**--CM-Sephadex— TSKgelG2000SW*--ConA-SepharoseCL6B ConA+ (desorption of >95% with 0,3M MM) At least 3 Asn-glycans (2kDa) PNGaseF+ (32—30kDa) [399] Rat small intestine mucosal brush-border Mb Retinyl ester hydrolase 130kDa (SDS-PAGE, for reduced protein) identical to phospholipaseB, solubilized with Brij35 ConA-Sepharose**—GlasPac-TSK-DEAE-5PW* ConA+ (desorption with 1,2M MM in the presence of OG; 47%, 6,5-f) [400] 3.1.1.6. Acetyl esterase Solanum tuberosum cv.Kennebec (potato tubers)

Enzyme of patatin SephadexG50—DEAEC—ConA-Sepharose ConA- (36%), ConA+ (desorption of 54% with 20mM MG) [401] 3.1.1.(6). Acetylesterase-like Schizophyllum commune ATCC 38548 (fungus) Acetylxylan esterase 18kDa (Superose12*) or 31kDa (SDS-PAGE) SephadexG25—DEAE-Sephadex— Q-Sepharose*—Phenyl-Sepharose**, ConA-Sepharose ConA- [402]

3.1.1.7. Acetylcholinesterase

Higher plant Synadenium grantii Hook (latex) Dimeric enzyme 70kDa, forms pI 5,0+5,2+5,4 CM-Sephadex—SephadexG200—PE-cellulose Carbohydr.: 14% including Ara(49-67% of total carbohydr.), Xyl(27-34%), Gal(4-7%), Glc(2-9%) [403]

Insects Drosophila melanogaster Canton-S (fruit fly, adult) Acetylcholinesterase of crude extract ConA-Sepharose ConA+ (almost complete binding),ConA- (in the presence of 0,5M MG) [406] Spodoptera frugiperda IPLB (Sf9,Sf21 cell lines; CF): Drosophila melanogaster acetyl- cholinesterase expressed in S.frugiperda cells using recombinant baculoviruses Enzyme 55+16kDa (SDS-PAGE), 70kDa (for precursor) ConA-Sepharose—Trimethylammonium-6- ConA (separation of hexylamine-sorbent—N-Methylacridinium- nonglycosylated forms sorbent 70kDa) [406a]

Other invertebrates Brugia malayi (filarial parasite, adult females) Monomeric secreted acetylcholinesterase forms 100 and 200kDa Cu2+-chelating-Sepharose**--ConA- ConA+ (binding of 95%; Sepharose desorption with 0,4M MM; 15%, 1,9-f) [404] Ciona intestinalis L.(ascidian larvae) Monomeric acetylcholinesterase 65-70kDa ConA-Sepharose4B—mCarboxyphenyl-methyl- ConA+(binding >95%; ammonium-iodide-Sepharose4B** desorption with 1M MM; 55-70%, 3-5-f) [405] Hirudo medicinalis (medicinal leech) Mb-bound acetylcholinesterase forms SS (66kDa [SephadexG200], or 30kDa [SDS-PAGE]), and DS (130kDa [SephadexG200], or 66kDa

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[SDS-PAGE]) solubilized with TX100 Edrophonium-Sepharose—ConA-Sepharose ConA+(desorption with 0,3M MG) [407] Fishes Electrophorus electricus (electric eel, electric organ) Acetylcholinesterase forms 1,900 and 310 kDa (SU 68+50+23,5kDa) associated with peptidase activity mAminophenyl-trimethylammonium-chloride- Sepharose**, Edrophonium-Sepharose, ConA-Sepharose ConA+ (binding of 80%; desorption by 1M MG with 0,1% TX100) [408] AcetylcholinesteraseG 260kDa ConA-Sepharose ConA+ (complete binding; desorption >75% with 0,5M MM; 10-f) [409] Acetylcholinesterase-D (18.4S) Lectin-agarose ConA+ (complete binding; desorption of 30% with 0,2M MM or MG), LCA+, WGA+, RCA+, Lectin- (around 60%) Carbohydr.: Man, Gal, GlcN, NeuAc [410]

Petromyzon marinus (lamprey skeletal muscle) Globular acetylcholinesteraseG4 (9,6S), and assymmetric form A12 (16S) ConA-Sepharose ConA+ (binding of 90%), ConA- (in the presence of 0,5M Glc) [411] Pleuronectes platessa (plaice muscle) Acetylcholinesterase solubilized with TX100 ConA-Sepharose ConA+ (complete binding; desorption with 0,5M MM; >75%, approx.10-f) [409] Amphibia Rana pipiens Schreiber (frog intact skeletal muscle) Surface acetylcholinesterase Inhibition with ConA (0,5mg/ml) ConA+ (20%) [412]

Mammals Bovine erythrocyte ghost Mb Acetylcholinesterase 145kDa (SDS-PAGE) or 77kDa (SDS-PAGE, for reduced protein) associated with lipid, solubilized with TX100 ConA-Sepharose ConA+ (complete binding without desorption by 0,3M MM), ConA- (in the presence of 0,3M MM) Acetylcholinesterase treated with

phosphatidyl inositol-specific phospholipaseC ConA-Sepharose ConA+ (binding and complete desorption with 0,3M MM) [413] Bovine different cell-type Mb Mb-bound acetylcholinesterase monomeric G1 (superior cervical ganglion), dimeric G2 (erythrocytes, lymphocytes, skeletal muscle), tetrameric G4 (Caudate nucleus, skeletal muscle, superior cervical ganglion), solubilized with 1-2% TX100

Bovine caudate nucleus Enzyme Lectin-Ultrogel ConA+(majority of native or Sialidase-treated enzyme),WGA+(native enzyme),UEA-I- Enzyme G4 Complex with lectin (sedimentation in ConA+ or WGA+ (complete sucrose gradient) binding), LCA+, UEA-I-

Bovine erythrocytes Enzyme Lectin-Ultrogel ConA-, WGA-, UEA-I-

Enzyme G2 Complex with lectin ConA-, LCA-, WGA-, UEA-I-

Bovine lymphocytes Enzyme Lectin-Ultrogel WGA-, LCA or UEA-I- (most of enzyme), LCA or UEA-I+ (desorption with 0,2M Man or L-Fuc, respectively), ConA+ (complete binding; desorption with 0,2M Man) Enzyme G2 Complex with lectin, WGA-, ConA+ (complete Inhibition in complex binding), LCA+, LTA+ (partial binding,inhibition of 35%), UEA-I+ (partial binding, inhibition of 70%, without inhibition in case of ConA-treated lymphocytes) Bovine skeletal muscle Enzymes G2, G4 Complex with lectin ConA+ or WGA+(partial), UEA-I-

Bovine superior cervical ganglion enzymes G1, G4 Complex with lectin ConA+ (complete for G1,

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partial for G4), LCA+ (partial for G4), WGA+ (complete for G4), WGA- (for G1), UEA-I- [414] Human erythrocyte Mb Acetylcholinesterase solubilized with 0,7% TX100 ConA-Sepharose4B, WGA-Sepharose6MB ConA+ (binding of 51%; desorption of 19% with 0,45M MG), WGA+ (binding of 73%; desorption of up to 68% with 0,45M GlcNAc; 59%, 19-f; separation from hemoglobin) Precipitation with lectins ConA-, LCA-, WGA- (for Sialidase-treated enzyme), PHA+ (40% at 15 mg/ml of PHA), WGA+ (90% at 5mg/ml of WGA) Carbohydr.: Sia C.perfringens sialidase+ [415]

Acetylcholinesterase solubilized with 2% TX100 Lectin-Ultrogel ConA+, WGA+ [414] ConA-Sepharose ConA+ (binding of 90%; desorption with 0,5M MM) [409] Human liver Monomeric acetylcholinesterase 68kDa ConA-Sepharose**--Sepharose4B ConA+ (desorption with 0,5M MG; 51%, 10-f) [416] Human serum Monomeric acetylcholinesterase 68kDa DEAEC**--ConA-Sepharose—Procainamide- ConA+ (desorption with Sepharose 50mM MG at pH 5,0; 81%, 3,5-f) [416] Rabbit erythrocyte Mb Mb-bound acetylcholinesterase of normal (I) or hypercholesterolemic (II) animals Inhibition with lectin SBA+ (20-65%, for I; 10-55%, for II) [417] Rabbit skeletal muscle sarcoplasmic reticulum and T-tubuli Globular monomeric, tetrameric, and assymetric acetylcholinesterase of fractions S1 and S2 solubilized with TX100 and separated by sucrose gradient Precipitation with lectins (1mg/ml) ConA2+, LCA2+, DBA+, WGA+, RCA-I+, VVA-B4+, PNA-, UEA-I- Asn-glycans C.perfringens sialidase- [418]

Rat brain Purified acetylcholinesterase forms pI 5,04+5,10+5,21+5,30+5,42+5,51 LCA-Sepharose LCA+ (binding >90% for

all forms; desorption of 30-40% with 0,25M MG) Carbohydr. Sialidase- [419]

Acetylcholinesterase forms fractionated with ammonium sulfate (10-40% saturation) LCA-Sepharose—WGA-Sepharose— LCA-, LCA+WGA-RCA-I-, RCA-I-Sepharose LCA+WGA+RCA-I-, LCA+WGA-RCA-I+, LCA+WGA+RCA-I+ (desorption with 0,12M MM,GlcNAc, or Gal, for LCA, WGA, or RCA-I, respectively) [420] Monomeric acetylcholinesterase 70kDa ConA-Sepharose—Dimethylaminoethyl- ConA+(desorption with benzoate-Sepharose sugar) [421]

Tetrameric acetylcholinesterase (10S) Ligatin-Affigel10, BiogelP100, Ligatin+ (desorption of DEAEC** 47% with 40mM Man-6-P or Glc-1-P; decreased binding of alkaline phosphatase- or EndoH- treated enzyme, by 30 or 60%, respectively) Asn-glycans of oligoMan-phosphorylated EndoH+ type [422]

Rat muscle Assymmetric collagen-tailed acetylcholin- esterase forms (I) of neuromuscular junctions, and globular forms (II) of more widely distribution + Lectin-agarose, ConA-agarose—VVA-B4- ConA (binding of 60-90% agarose for I and II; desorption with 0,5M MM), DBA+ or + VVA-B4 (binding of 20- 25%; desorption of I with 0,2M GalNAc; separation from II) [423] Sheep brain basal ganglia Acetylcholinesterase 470kDa, SU 50+23.5 kDa, associated with peptidase activity mAminophenyl-trimethylammonium-chloride- Sepharose**, Edrophonium-Sepharose, ConA-Sepharose ConA+ (desorption by 1M MG with 0,1% TX100) [408] 3.1.1.8. Pseudocholinesterase

Fish Acanthuras dussumieri (surgeonfish tissue) Globular tetrameric pseudocholinesterase sialoform 250kDa (11,5S) QAE-Sephadex—ConA-Sepharose** ConA+ (complete

14

binding;washing with 0,1M MG; desorption with 30mM MM; 91%, 28-f) Precipitation with ConA (0,3mg/ml) ConA+(65%) [424]

Mammals Human serum Monomeric pseudocholinesterase 90kDa associated with peptidase activity ConA-Sepharose ConA+ (complete binding; desorption >90% with 0,5M MG) [425] Monomeric 90kDa associated with arylacylamidase activity RCA-I-Sepharose RCA-I+ (complete binding; desorption of 97% with 0,5M Lac) Chymotrypsinized Gp 20kDa with butyryl- cholinesterase and arylacylamidase activities RCA-I-Sepharose RCA-I+ Carbohydr.: 8.8% (including Man, Gal), for 90kDa; 7% (including Man), for 20kDa [426]

Human sera of patients with liver cirrhosis and chronic hepatitis Cholinesterase forms with affinity to lectins AAL+ [426a]

3.1.1.11.

Lower plants Aspergillus niger (CF) or Saccharomyces cerevisiae W3124 (CF): Fungal Aspergillus aculeatus pectin methyl esterase expressed in yeast or other fungal cultural cells Enzyme-I 138aar (36,2 kDa, pI 3,8) SDS-PAGE—Blot—Lectin DSA-, GNA-, PNA- Carbohydr.(M/M): Gal(1),GlcN(3),Man(10); Thr244-glycan; Asn156,227-potential glycans [427]

Phytophthora infestans (fungal CF)

Monomeric pectinesterase forms I (45-48 kDa), and II (35-40kDa) DEAEC—CMC—Chromatofoc—SephacrylS200 —ConA-Sepharose ConA+ (form I) [428]

Higher plants Actinidia chinensis cv.Hayward (kiwi fruits) Monomeric pectinmethylesterase 57kDa, pI 7,3 DEAEC—Heparin-Sepharose—Superose12**--

ConA-Sepharose* ConA+ (desorption with gradient 0-80mM Glc; desorption of forms at 25th and 28th min) [429] Citrus sp. (mandarin orange fruits) Monomeric pectinmethylesterase 37kDa, pI>8,65 DEAEC—Heparin-Sepharose**--Superose12* Carbohydr. DIG-kit+ (Boehringer, Germany) [430] Glycine max L. (cells in culture) Homogeneous cell-wall pectinmethyl- esterase (SDS-PAGE) Dot-Blot—Lectin (in combination with ConA+ alpha-1,3-Fuc- or beta-1,2-Xyl-specific Ab) GNA+ Glycans: Fuc of glycans similar to that SBA+ 2+ of honey bee phospholipaseA2; Xyl of TPA glycans similar to that of horseradish peroxidase [431]

Lycopersicon esculentum cv.VF 145B-7879 (tomato fruit cell walls) Purified pectinmethylesterase ConA-Sepharose4B ConA+ (separation from polygalacturonase) [432] 3.1.1.13. Sterol esterase

Human foreskin fibroblast CF Monomeric cholesterol esterase associated with acidtriacylglycerol lipase, 47-49kDa ConA-agarose ConA+ (desorption with MM) Asn-glycans of oligoMan type EndoH+ [374]

Human liver lysosomes Monomeric cholesterol esterase associated with acid lipase, 59-69kDa ConA-Sepharose—CMC—MonoS* ConA+ (desorption with 0,4M MM) [376] Sterol esterase/acid lipase 56+41kDa (SDS-PAGE, 41kDa as catalytic SU) solubilized with TX-100 ConA-Sepharose**—CM-Sepharose— ConA+ (desorption with Phenyl-Superose*—MonoS*—Superose12* 0,37M MM; 50%, 4,9-f) Asn-glycans of oligoMan type: 4kDa in EndoH+ (41—37kDa) 41 kDa [433]

Rabbit arterial tissue Lysosomal cholesterol esterase ConA-Sepharose ConA+ (desorption of 76% with with 0,1M MM; separation from the bound lipoproteins) [434] 3.1.1.14. Chlorophyllase Phaeodactylum tricornutum (diatomaceous alga)

16

Mb-bound monomeric 3-4 forms PAS+ (all forms) SephadexG100—ConA-Sepharose4B ConA+ (binding of 95%; desorption of 75% with 0,2M MM) [435] 3.1.1.17. Gluconolactonase Fungal cells Lactonase 70kDa, pI 5,2-5,7, of commercial cellulase complex Novozym 188 (Novo, Kopenhagen) DEAEC—SepharoseCL6B—ConA-agarose— ConA+ (complete binding; Chromatofoc—SephacrylS200—IEF desorption with 0,5M MM) Carbohydr.: 40% [436]

3.1.1.23. Acylglycerol lipase

Higher plant Solanum tuberosum cv.Kennebec, cv. Russet Burbank Laurate-esterase of patatin SephadexG50—DEAEC—ConA-Sepharose** ConA- (3%), ConA+ (desorption with 20mM MG; 86%, 2,0-f) [401]

ConA- (27%), ConA+ binding of 73%; desorption of 20% with 20mM MG, and additional 24% with 0,3M MG) [396] Mammal Rat lung cell cytosol Monomeric caprylate-esterase 77kDa, pI 4.1 DEAEC**—HA—Octyl-Sepharose—ConA- ConA+(desortion with Sepharose 0,1M MM; 57%, 1,7-f) [437] 3.1.1.32. Phospholipase A1

Human plasma + PhospholipaseA1 64kDa (SDS-PAGE), PAS pI 4,1 Heparin-Sepharose**--ConA-Sepharose ConA+ (desorption with 1M MG; 45%, 2,2-f) [378] Rat kidney cortex lysosomes PhospholipaseA1 forms 70 (SU 34kDa) and 25kDa (HA—ConA-Sepharose)**--Chromatofoc— ConA+ (desorption by Sephadex-G150 0,5M MM with 5mM EDTA; separation from inhibitors), HA—ConA+ (230%, 254-f) [438] Rat liver lysosomes PhospholipaseA1 associated with lysophospholipase, forms 90kDa (pI 5,2), 34kDa (pI 5,2), 15kDa (pI 5,2), 44kDa (pI<4), 24kDa (pI<4) HA—DEAEC—ConA-Sepharose**--Chromatofoc ConA+ (desorption with (pI 5)—Chromatofoc(pI 4)— UltrogelAcA44 0,5M MM; 40%, 4,6-f) [398]

ConA-Sepharose—HA—DEAEC (or Chromato- ConA+ (complete foc) binding; desorption of 40-60% with 0,5M MM) [398] Monomeric phospholipaseA1 56kDa ConA-Sepharose**—Phenyl-Sepharose— ConA+ (desorption with Sephadex- -Sephadex 0,3M MM; 42%, 2,3-f; reactivation of enzyme with 10mM CaCl2) [439] Phospholipase-A1 ConA-Sepharose**--Chromatofoc— ConA+ (desorption with Fractogel-TSK-HW55S 0,5M MM; separation from inhibitors; 156%, 165-f) [440] ConA-Sepharose ConA+ (recovery 90%, approx. 1,000-f) [441] (Orange-agarose—Red-agarose)—(ConA- ConA+ (desorption with Sepharose—Phenyl-Sepharose) 0,5M MM) [442]

(3.1.1.32.) Protein inhibitors of phospholipase A1 Rat liver lysosomes Intralysosomal GP as inhibitors of endogenic phospholipaseA1 [441]

3.1.1.34. Lipoprotein lipase

Bird Hen adipose tissue and adipocytes in culture Monomeric lipoprotein lipase 59,7, or 52kDa (from tunicamycin-treated cells) ConA-Sepharose ConA+ (separation of EndoF-treated glycans; desorption with 0,2M MG followed by 0,2M MM) Carbohydr.moiety: 7,7kDa, 2 Asn-glycans of complex type, and 1 EndoH+ (elimination of Asn-glycan of oligoMan type, Asn45- sulfate groups), EndoF+ GlcNAc-sulfate [443]

Mammals Bovine adipocytes 3T3-L1 (CF) Monomeric lipoprotein lipase 57 or 48kDa (from tunicamycin-treated cells) Carbohydr.: 9kDa, 3 Asn-glycans [444]

Bovine brain microsomes Mb-bound lipoprotein lipase solubilized with 0.2% TX100 ConA-Sepharose ConA+ (complete binding; desorption with 0,5M MM; 45-50%, 3-5-f) [445] Guinea pig adipocytes Lipoprotein lipase of cell extract Lectin-agarose ConA+ (intact or Sialidase-treated form),

18

RCA-I+ (binding and desorption of 40% with 0,2M Gal, intact form; binding of form EndoH-Sialidase+) [446] Human postheparin plasma Monomeric lipoprotein lipase 58kDa Heparin-Sepharose**—Heparin-Sepharose— HA—ConA-Sepharose ConA+ (complete binding; desorption by 1M MG with 10mM CaCl2; 71%, 85-f) [447] SDS-PAGE—Blot—ConA—Peroxidase ConA+ [448]

Rat newborn heart cells in culture Extracellular monomeric lipoprotein lipase 56 or 49kDa (from tunicamycin- treated cells) Carbohydr.: 7kDa [449]

3.1.1.53. Sialate O-acetylesterase

Rat liver cell Golgi vesicular lumens Mb-bound (I) and cytosolic (II) sialate serine esterase ConA-Sepharose ConA+ (for I;separation from II) [450] Rat liver microsomal Mb 9-O-Acetylated sialic acid-specific esterase 61,5kDa (SU 36+30kDa) DEAEC—ConA-Sepharose**—Sephacryl- ConA+(desorption with S200—Procion-Red-agarose 0,1M MM; 68%, 31-f; separation from cytosolic form) Asn-glycans of oligoMan type and EndoH+ complex type EndoF+ [451]

Rat liver and FAO hepatoma cell microsomal Mb Sialic acid-specific O-acetylesterase precursor 65kDa and mature form 58kDa, SU 38+28kDa Asn-glycans of complex type: EndoH- 12kDa in 65kDa PNGaseF+ (65—53kDa) [452]

3.1.1.59. Juvenile hormone esterase Trichoplusia ni (moth, hemolymph and bodies at the last larval star) Monomeric enzyme 64kDa SDS-PAGE—Blot—ConA ConA+, ConA- (for EndoF- treated enzyme) Asn-glycans EndoF+ [453]

Juvenile hormone esterase forms pI 5,3 and 5,5 ConA-Sepharose4B ConA+ (binding of 88-92%, or 79-86%, for normal or juvenile hormone analog-

treated larvae, respectively) [454] 3.1.1.-. Colletotrichum lagenarum (Pass.) Ell and Halst. race 1 (fungal cucurbitanthracnose pathogen, CF) Monomeric cutinase "a", 60kDa, acid pI DEAEC—Octyl-Sepharose—Octyl-Sepharose— ConA-Sepharose4B ConA+ (desorption with 0,3M MM) [455] 3.1.3.1. Alkaline phosphatase

Lower plants Dictyostelium discoideum (slime mold plasma Mb at different amoebial stages) Enzyme solubilized with Na-DOC ConA-Sepharose ConA+ (binding of 92, 77, or 57%, for vegetative, preaggre- gated, or aggregated cells, respectively; desorption with 0,1M MG) [456] Vegetative cell Mb-bound alkaline phosphatase associated with 5'- nucleotidase, solubilized with TX100 ConA-Sepharose**—DEAE-Sephacel ConA+ (desorption with 0,2M MM; 74%, 8,5-f) [457] D. discoideum strain NC4 Mb-bound alkaline phosphatase (>300kDa) solubilized with TX100 ConA-Sepharose ConA+ (desorption with 1M MM; separation from neutral phosphatase and histon-phosphatase) [458] Saccharomyces cerevisiae 1710 (yeast protoplast vacuoles) Alkaline phosphatase 130kDa, forms I-III, SU 66+92+46kDa DEAEC—Phenyl-agarose—4(4-aminophenyl- azo)phenylarsonate-agarose—DEAEC— BiogelP200—ConA-Sepharose ConA+ (for I and II,SU 92+46kDa; desorption with 25mM MM) Precipitation with ConA ConA+ (96 or 54%, for native or tunicamycin- modified I), ConA+ (99 or 63%, for II),ConA+ (9 or 4%, for III) Carbohydr.: 8 or 15%, in 92 or 46kDa, EndoH+ (92 and 46kDa) respectively; Asn-glycans of oligoMan-type [459]

Invertebrates Bombyx mori (silkwarm, 5th instar larvae, midgut)

20

Alkaline phosphatase Mb-bound form 68kDa, and cytosolic soluble form DEAE-Sephacel—ConA-Sepharose**-- ConA+ (desorption with UltrogelAcA34 15mM MM using sugar gradient 0-25mM; 98%, 15-f, for 68kDa) [460] Alkaline phosphatase Mb-bound form 58kDa(SDS-PAGE),and soluble form 61 kDa(SDS-PAGE) SDS-PAGE—ConA-peroxidase ConA+ Carbohydr.: 2.1 or 8.4%, for 58 or 61 kDa, respectively Carbohydr.composition (M/M): hexoses (5 or 20, for 58 or 61kDa, respectively), GlcNAc (1 or 2), GalNAc (1 or 6) Asn-glycans: asialylated afucosylated biantennary of complex type (58kDa), oligoMan type (61kDa) Lectin-agarose (combined ConA+WGA-, ConA+WGA+, - - - + chromatography) ConA PHA-E4 , ConA PHA-E4 (separation of glycans) [461] Ceratitis capitata (mediterranean fruit fly, integument) Alkaline phosphatase major form 180kDa, and minor form 900kDa (associated with chitin), solubilized with TX100 ConA-Sepharose—SephadexG200— ConA+ Sepharose6B [462]

Echinococcus mutilocularis (helminth, metacestodes) Alkaline phosphatase 240kDa, SU 80kDa, pI 4,7+4,8 ConA-Sepharose ConA+ (desorption with MM; 16%, 10-f) [463] Pandalus borealis (shrimp hepatopancreas) Alkaline phosphatase 155kDa, SU 70kDa Q-Sepharose(*)**--ConA-Sepharose— ConA+ (desorption with Procion-Red-Sepharose-HE3B 10mM MG; 78%, 3,8-f) [464] Penaeus japonicus bate (shrimp hepatopancreas Mb) Monomeric alkaline phosphatase 55kDa DEAEC—ConA-Sepharose—Ultrogel- ConA+ (desorption with AcA34—Prepar.PAGE** MM; 47%, 4,7-f) Asn-glycans: 12kDa in 55kDa EndoF+ (55—33kDa) [465]

Penaeus monodon (shrimp hepatopancreas) Alkaline phosphatase forms B1 and B2 (200kDa, SU 31+43kDa), and form A (300 kDa, SU 31kDa) DEAEC—SephacrylS200**--ConA- ConA+ (desorption of A Sepharose with 30mM MM; 70%, 2,6-f; desorption of B1 and next

B2 in the presence of 30 mM MM with increasing ionic power; 23%, 6,0-f, for B1; 65%, 6,2-f, for B2) [466] Bird Hen intestine Commercial alkaline phosphatase (SISCO Res.Lab.,India) (ConA-crosslinked with glutaraldehyde)- ConA+ (desorption of DEAEC, ConA-agarose (Hygro Chemicals, 55-60% with 0,1M Man) India) [467]

Mammals Bovine brain Alkaline phosphatase solubilized with 0,25% Na-DOC ConA-Sepharose ConA+ (desorption by 0,5 M MG with 0,1% Na-DOC; approx. 4-f) [468] Bovine intestine brush border Mb Mb-bound alkaline phosphatase Affinity to lectin-coated bottoms of ConA+ ELISA plate [469]

Purified alkaline phosphatase non-enzymically glucosylated (in the presence of 5mM Glc, 24 h, 37oC), neoGP with 23%-decreased activity [7]

Small intestinal alkaline phosphatase forms pI 4,5-4,9, and pI<3.7 IEF-PAG—Blot—WGA-peroxidase WGA+ [470]

Bovine mammary gland microsomal Mb Dimeric alkaline phosphatase 79.5 kDa, 30% carbohydr. SephadexG200—ConA-Sepharose** ConA+ (washing with 50mM MM; desorption with 0,5M MM; 90%, 30-f) [471] Bovine milk Alkaline phosphatase SephadexG200**—DEAEC—SephadexG200 —ConA-Sepharose4B ConA+ (desorption with 0,25M MM; 80%, 2,2-f) [472] Bovine neutrophils (polymorphonuclear) Monomeric alkaline phosphatase 165kDa Sephadex-G200**--DEAEC—SephadexG200 —ConA-Sepharose—SephadexG200 ConA+ (binding of 90%; desorption with 30-40 mM MM using gradient 0-0,2M sugar; 50%, 2,0-f) [473] Bovine thyroid cell plasma Mb Alkaline phosphatase solubilized with 1.5% Na-DOC LCA-Sepharose4B LCA- (separation from 75% 5'-nucleotidase)

22

Inhibition of Mb-bound enzyme with ConA ConA+ (3%) [474]

Dog liver of animal treated with drugs Alkaline phosphatase Mb-bound form 105 kDa (145 or 290kDa, for phospholipaseC- or sialidase-treated enzyme, respectively), soluble form 130kDa (270 kDa for sialidase-treated enzyme), and 136kDa solubilized with n-butanol SephadexG200—DEAEC, ConA-Sepharose ConA+ (complete binding of 145 and 290kDa; desorption of 17 or 77% with 10mM or 0,5M MM, respectively, for Mb- bound forms; desorption of 34 and next 62% with 10mM MM followed by 0,5M MM, for soluble forms) Carbohydr.: Sia Sialidase+ [475]

Dog serum (beagle dog) Alkaline phosphatase forms: intestinal- Like (I), hepatic-like (II),and bone- Like (III), pI 3,1-5,4 SephadexG200—DEAE-Sephadex—ConA- ConA- (52%, for pI 3,3- Sepharose4B 4,0), ConA+ (desorption of 30% with 50mM MM,for I and II, pI 4,0-5,4, 3,1; desorption of next 19% with 0,5M MM, for III) [476] Alkaline phosphatase (SU 75-78kDa) with high affinity to ConA SephadexG200—DEAE-Sephadex—ConA- ConA+ (washing with 50 Sepharose4B** mM MM; desorption with 0,5M MM; 47%, 3,7-f; or 52%, 3,4-f) [477] Purified enzyme LAE in agarose WGA+ (35 or 43%, for male or female sialoforms, respectively) [477] Dog serum of animals treated with phenobarbital and brovanexine Alkaline phosphatase 270kDa treated with phospholipaseC and sialidase ConA-Sepharose ConA+ (complete binding; desorption of 34% and next 62% with 10mM MM followed by 0,5M MM) [475] Human amnion cells FL (Mb) Purified alkaline phosphatase Kasahara isozyme like intestinal type enzyme AAL-Sepharose—BiogelP4—ConA- AAL+, ConA+ (separation Sepharose of glycans) [478] Carbohydr.: Sia Sialidase+ Asn-glycans: fucosylated bi- and

polyantennary

Human bone Alkaline phosphatase of extract Precipitation with lectins ConA+ (51 or 47%, for native or Sialidase- treated enzyme, respect- ively), WGA+ (78 or 24%), PHA+ (90 or 69%), RCA-I+ (96 or 91%), SBA+ (2 or 38%), PNA-, UEA-I- [210] Monomeric alkaline phosphatase forms 160 and 165kDa LAE in agarose WGA+ Carbohydr.: Sia [479]

Human duodedenum Alkaline phosphatase 148kDa,forms pI 4,3 and 6,3 ConA-Sepharose ConA- (>50%), ConA+ (desorption with 10mM MM) [480] Alkaline phosphatase 140kDa(SDS-PAGE) LAE in agarose WGA+ [479]

Alkaline phosphatase of extract Precipitation with lectins ConA+ (52 or 59%, for native or Sialidase- treated enzyme, respect- ively), WGA+(87 or 27%), PHA+ (87 or 64%), RCA-I+ (90 or 94%), SBA+ (24 or 47%), PNA-, UEA-I- [210] Human erythrocytes Monomeric alkaline phosphatase 17-18kDa DEAE-Sephadex**--pAmino-benzylphospho- nate(ABPA)-Sepharose—pABPA-Sepharose —ConA-Sepharose ConA- (for all 6 isozymes) [481] Human ileal tissue Alkaline phosphatase 134kDa, forms pI 6,1 and 6,8 ConA-Sepharose ConA+ (desorption with 10mM MM) LAE in agarose WGA+ [480]

Alkaline phosphatase 130kDa(SDS-PAGE) LAE in agarose WGA+ [479]

Human intestine Alkaline phosphatase 140kDa ConA-Sepharose-4B**— Tyramin-As- ConA+ (desorption with Sepharose 0,3M MM; 46%, 68-f)

Activation with ConA (20nM) or PHA ConA2+ (75-85%), (2mg/ml) PHA-P+> PHA-M+

24

Carbohydr.composition [482,483] Alkaline phosphatase of extract Precipitation with lectins ConA+ (41 or 44%, for native or Sialidase- treated enzyme, respectively), WGA+ (53 or 8%), PHA+ (66 or 26%), RCA-I+ (55 or 62%), SBA-, PNA-, UEA-I- [210] Soluble and particulate alkaline phosphatase forms from brush-border ConA-Sepharose ConA+ (43, 2 or 54%, for total, particulate, or soluble activity, respectively; desorption with 0,2M MM) [484] Human kidney, cortex and medulla Alkaline phospholipase of kidney extract Precipitation with lectins ConA+ (39 or 32%, for native or Sialidase- treated enzyme, respectively), WGA+ (80 or 13%), PHA+ (93 or 73%), RCA-I+ (93 or 95%), SBA+ (8 or 59%), PNA-, UEA-I- [210] Alkaline phosphatase cortex form 380kDa,and medullar form 360kDa ConA-Sepharose, ConA+ and WGA+ WGA-affine electrophoresis (LAE) (simillarity between in agarose cortex and duodenum forms,or between medullar and ileal forms) Asn-glycans: relatively high content of polyantennary of complex type (cortex enzyme),or hybrid type (medullar enzyme) [485]

Alkaline phosphatase forms 165 and 125kDa(SDS-PAGE) LAE in agarose WGA+ [479]

Human leukocyte (polymorphonuclear) Alkaline phosphatase DEAE-SepharoseCL6B—Lectin-Sepharose(or ConA+ (binding of 80%; Cibacron-Red-Sepharose4B desorption with 0,25M MM; 80%, 2-f), WGA+ [486] Human lung from patient with cancerous pleuritis Alkaline phosphatase of extract Lectin-agarose (combined chromatography) ConA-PHA-E+ (21 or 2% for soluble or Mb-bound enzyme, respectively), ConA-PHA-E+ (5 or 0%), ConA+PSA- (7 or 39%), ConA+PSA+ (50 or 10%),

ConA2+WGA- (16 or 15%), ConA2+WGA+ (1 or 26%) [487] Human liver Alkaline phosphatase of extract Lectin-agarose ConA+ and LCA+ (partial binding), WGA-, HPA- [488] ConA+ (complete binding; desorption of 67 and next 33% with 10mM and 0,5M MM, respectively), E-PHA- [489]

Sequential lectin chromatography ConA+PSA- (30%), ConA+PSA+ (37%, desorption with 10mM MM or 0,2M MM, respectively), ConA2+WGA+ (33%; complete binding to WGA; desorption with 0,5 M MM or 0,1M GlcNAc) [489] Precipitation with lectins ConA+ (89 or 79%,for native or Sialidase- treated enzyme,respect- ively), WGA+ (87 or 1%), PHA+ (69 or 61%), RCA-I+ (96 or 84%), SBA-, PNA-, UEA-I- [210]

-pr -G200 ConA+ (separation from inhibitors; solubilization with 0,7M MG) [490] Dimeric alkaline phosphatase 150kDa DE-nugel—ConA-Sepharose—ConA- ConA+ Sepharose—MonoQ*—Phenyl- sorbent(*)**—C8-sorbent* Asn-glycans: 25kDa in 75kDa EndoF+(75—50kDa) [13]

Alkaline phosphatase 175kDa Inhibition with ConA (40nM) ConA+(8-15%) Carbohydr.: Sia Sialidase+ [482]

Human malignant mesothelioma of omentum Alkaline phosphatase of extract LAE on cellulose acetate plate WGA+ (for native or subtilisin-treated enzyme) Asn-glycans of complex type EndoF+ [489]

Human milk Alkaline phosphatase 160kDa DEAEC—ConA-Sepharose ConA+ (desorption with MM) Carbohydr.: Sia Sialidase+ [491]

26

Human ovarian epithelial tumors Alkaline phosphatase, SU 63,5kDa Ab-sorbent—SephadexG200—Phenyl- Sepharose (or ConA-Sepharose) ConA+ (binding of 82%) Carbohydr.: different contents of Sia Sialidase+ (for ConA- in ConA- and ConA+ forms and ConA+ forms) [492]

Human placenta Alkaline phosphatase phenotypes FI(134kDa, Caucasian),SS(128kDa, Japanese), and FF (128kDa, Negroid) Lectin-agarose (sequential chromatography) ConA+ (complete binding of all forms), ConA+PSA- (36, 19, or 13%, for FI, SS, FF, respectively), ConA+PSA+ (26, 27, or 48%), ConA2+WGA- (18, 41, or 24%), ConA2+WGA+ (21,13,or 16%) Asn-glycans in FI and FF (6 kDa in FI) EndoF+ (FI, FF), Sialidase- [493] Alkaline phosphatase of extract Lectin-agarose (sequential chromatography) ConA-PHA-E- (0%), ConA-PHA-E+ (0%), ConA+PSA- (13%), ConA+PSA+ (43%), ConA2+WGA- (28%), ConA2+WGA+ (16%) [487]

Lectin-agarose ConA+ or LCA+ (high affinity), LCA- (for a small part of enzyme), WGA+ (complete binding), HPA- [488] LAE on cellulose acetate plate WGA+ [489]

Crude alkaline phosphatase Type XVII (Sigma,USA), isozymes A and B DEAEC**--ConA-Sepharose ConA+ (73%, 2,2-f, for A; 53%, 6,8-f, for B; washing with 1mM MM; desorption with 50mM MM) [494] Alkaline phosphatase 160kDa DEAEC—ConA-Sepharose—SephacrylS200 ConA+ (desorption with MM) Carbohydr.: Sia Sialidase+ [491]

Alkaline phosphatase 130kDa LAE in agarose WGA+ [479]

Human primary hepatoma Soluble akaline phosphatase Precipitation with lectins ConA+ (61 or 75%, for native or Sialidase- treated enzyme,

respectively), WGA+ (93 or 28%), PHA+ (77 or 54%), RCA-I+ (93 or 100%), SBA- ,PNA-, UEA-I- [210]

LAE on acetate cellulose plate WGA+ (for subtilisin- or EndoF-treated enzyme) Asn-glycans; Sia in Ser/Thr-glycans EndoF+ [489,495] Purified Kasahara isozyme forms I-IV similar to intestinal type enzyme LAE in agarose ConA+ (complex with II; dissociation by 0,2M MM) [496] Human seminomas Alkaline phosphatase forms pI 4,3-4,6 Lectin-agarose (sequential chromatography) ConA-WGA- (19%), ConA-WGA+ (9%), ConA+WGA- (48%), ConA+WGA+ (23%), desorption with 10mM MM or 0.5M GlcNAc, for ConA or WGA, respect- ively) Carbohydr.: Sia Sialidase+ (pI [4,3-4,6]— [4,6-5,0]) [497] Human serum from healthy donors Bile-, bone-, and liver-like alkaline phosphatase forms (I, II, III, respectively) WGA-Sepharose4B WGA+ (for II), WGA- (for III) [498]

WGA-Silica* WGA+ (separation of 81% of III, and 92% of II, using gradient of GlcNAc) [499]

LAE in agarose WGA+ (heterogeneity of (488 donors [701]) I-III) [221,500-504] WGA+ (enzyme heterogen- eity depending on age, sex, drug-treatment) [505,506] Precipitation with lectins (108 donors, ConA+ (75-85%), SEL+, different commercial WGA, [1213]) WGA+ (46%) [221,507-509] CEA+ (50%) [510] WGA+ (80-95% of II) [509,511,512] LAE on cellulose acetate plate WGA+ [512] Alkaline phosphatase of children serum ConA-Sepharose4B, WGA-Sepharose6MB ConA- (10%), ConA+ (desorption of 84 and next 6% with 10mM MM followed by 0,5M MM), PSA- (41%), PSA+ (desorpt- ion of 59% with 0,2M MM),WGA+ (complete binding; desorption

28

with 0,2M GlcNAc), WGA- (35%, for Sialidase- treated enzyme), WGA+ (desorption of 38 and next 27% of Sialidase- treated enzyme with 0,2 M and next 0,5M GlcNAc) [513] Alkaline phosphatase of adult serum Lectin-Sepharose4B (sequential chromatog- ConA- (6%), ConA+ (desorption of 54% with 10mM MM), ConA2+WGA- (8%, desorption with 0,5M MM), ConA2+WGA+ (32%) [479]

Combined/ sequential precipitation LCA+ or/and WGA+ with lectins (87 adults, 72 childs of 6-15 years) (differentiating of II) [514]

Human serum of patients Alkaline phosphatase from healthy women in early postmenopause (age of 46-53 years) before and after receiving hormone therapy with estrogens for prevention of bone loss; from patients with hepatobiliary disease or liver metastasis upon stomach cancer Precipitation with WGA WGA+ [509]

Alkaline phosphatase from patients with bone or hepatobiliary diseases LAE WGA+ [512,515]

Alkaline phosphatase from patients with failing liver function LAE in Titan-agarose WGA+ (separation of (Helene Labs, Beaumont, TX, USA) II, III, and forms of III) [516]

Human skin blood vessels of aorta Alkaline phosphatase DEAEC—ConA-Sepharose—SephadexG150 ConA+ (desorption with 0,5M MG) Carbohydr.: Sia Sialidase+ [517]

Human testis Alkaline phosphatase forms 4,1, 5,0-5,2 Lectin-agarose (sequential chromatography) ConA-WGA- (17%), ConA-WGA+ (18%), ConA+WGA- (36%), ConA+WGA+ (29%) Carbohydr.: Sia Sialidase+ (pI 4,1—5,2) [497]

Human urine Alkaline phosphatase of healthy adults (liver-like monomeric enzyme 152kDa),and children (bone-like monomeric enzyme 165

kDa) Lectin-Sepharose (combined ConA- (8 or 10, for adult chromatography) or child, respectively), ConA+ (44 or 17%), ConA2+WGA- (12 or 0%), ConA2+WGA+(36 or 73%)[479,518] Pregnant women Monomeric alkaline phosphatase placental- like form 135kDa, bone form 165kDa, and kidney form Lectin-Sepharose4B (sequential ConA- (6%), ConA+ (24%; chromatography) desorption with 10mM MM), ConA2+WGA- (57%), ConA2+WGA+ (23%) [479] Patient with primery hepatoma Alkaline phosphatase monomeric major form 140-150kDa, and minor form 165kDa (SDS-PAGE) Lectin-Sepharose4B (sequential ConA- (8%), ConA+(17%; chromatography) desorption with 10mM MM), ConA2+WGA- (4%; desorption with 0,5M MM), ConA2+WGA+ (71%) [479,518] Patient with chronic nephritis Monomeric alkaline phosphatase forms 165, 152, and 130kDa (SDS-PAGE) Lectin-Sepharose4B (sequential ConA- (43%), ConA+ (26%), chromatography) ConA2+WGA- (8%), ConA2+WGA+ (23%) [479,518] Patient with acute nephritis Monomeric alkaline phosphatase forms 165, 152, and 125kDa (SDS-PAGE), 125kDa as kidney form Lectin-Sepharose4B (sequential ConA- (46%), ConA+ (17%), chromatography ConA2+WGA- (9%), ConA2+WGA+ (28%), ConA- or L-PHA-(for 125 kDa) [479,518] Psychiatric disease patient given daily 300mg chlorpromazine administration Lectin-Sepharose4B (sequential ConA- (9%), ConA+ (21%), chromatography) ConA2+WGA- (3%), ConA2+WGA+ (17%) [479,518] Murine uterus Alkaline phosphatase 205kDa solubilized with TX100 ConA-Sepharose**--DEAEC—SephacrylS200 ConA+ (desorption with — SephacrylS200 50mM MM; 65%, 10-f) [519] Porcine kidney Alkaline phosphatase ConA-Sepharose, Ab-sorbent** ConA+ [520]

30

Rabbit kidney Alkaline phosphatase with SU 72kDa (SDS- PAGE), as intestinal-like enzyme mAb-CH-Sepharose—Sepharose — SephacrylS200, ConA-Sepharose ConA+ (separation of 3 forms with total recovery >90%: unbound, eluted with 10mM MM,and then eluted by 0,5M MM) Carbohydr.: Sia Sialidase+ [521]

Rat bone Alkaline phosphatase Lectin-agarose (sequential ConA-PHA-L+ (7%), ConA+ chromatography) (85%), ConA+PSA- (6%), ConA+PSA+ (12%), ConA2+WGA+ (75%) [522] Rat duodenum Alkaline phosphatase Lectin-agarose (sequential ConA-PHA-L+ (46%), ConA+ chromatography) (106%), ConA+PSA- (40%), ConA+PSA+ (8%), ConA+WGA+ (6%) [522]

ConA-Sepharose4B ConA+ (complete binding; desorption of 72% and next 28% with 10mM MM followed by 0,5M MM) [523,524] Rat duodenal explants in culture Alkaline phosphatase ConA-Sepharose ConA- (23 or about 15%, for without or with swainsonine in culture, respectively), ConA+ (desorption of 54 or about 40% with 10mM MM, and next 23 or 60% by 0,5M MM, for without or with swainsonine in culture, respectively) [522] Rat hepatoma AH130 cell microsomes Alkaline phosphatase forms 160 and >500kDa (SephacrylS300),solubilized with n-butanol at pH 5,5 (160kDa) or pH 8,5 (> 500kDa) ConA-Sepharose**--SephacrylS300—HA ConA+ (desorption with 0,4M MG; 65%, 5,5-f) [525] Rat hepatoma JTC16 cells Alkaline phosphatase 290kDa, SU 72kDa DEAEC—SephadexG200—ConA-Sepharose— ConA+ (desorption with —PAGE** 50mM MM; 72%, 3,2-f) [526]

Rat kidney Alkaline phosphatase Lectin-agarose (sequential ConA-PHA-L+ (5%), ConA+ chromatography) (91%), ConA+PSA-(31%),

ConA+PSA+(47%), ConA2+WGA+(17%) [522] Rat liver Alkaline phosphatase Lectin-agarose (sequential ConA-PHA-L+ (3%), ConA+ chromatography) (92%), ConA+PSA+ (11%), ConA+PSA+ (41%), ConA2+WGA+ (45%) [522] ConA+PSA-, ConA+PSA+ [527] ConA-agarose ConA- (9%), ConA+ (desorption of 73 and next 19% with 0,1M MM followed by 0,3M MM) [528] Asn-glycans Lectin-agarose ConA+ (desorption of 41% and next 1% with 0,1M followed by 0,3M MM; for Sialidase-treated glycans; separation from unbound 58% glycans), PHA-E+ (for EndoH-treated glycans; retardation of poly- antennary glycans) [528] Rat serum Alkaline phosphatase Lectin-agarose (sequential ConA-PHA-L+ (2%), chromatography) ConA+(96%), ConA+PSA- (8%), ConA+PSA+ (28%), ConA2+WGA+ (62%) [522] Rat serum duodenal alkaline phosphatase form ConA-Sepharose ConA+ [529]

Rat serum of streptozotocin-induced diabetic animals (28 days after inducing)

Elevated level of alkaline phosphatase ileal form, and other forms ConA-Sepharose4B ConA- (26%), ConA+ (desorption of 65% with 10mM MG, and next 8% by 0,5M MG) [524] Rat small intestine Alkaline phosphatase forms pI 8,3-9,5, 4,5-4,9 IEF-PAG—Blot—WGA-peroxidase WGA+ [470]

Intestine ileal alkaline phosphatase ConA-Sepharose4B ConA- (15%), ConA+ (desorption of 73% with 10mM MG, and next 12% by 0,5M MG) [523,524] Lectin-agarose (sequential ConA-PHA-L+(17%), ConA+

32 chromatography) (103%), ConA+PSA- (47%), ConA+PSA+ (26%), ConA2+WGA+ (10%) [522] Proximal (I) and distal (II) small intestinal brush-border Mb-bound alkaline phosphatase forms Lectin-Sepharose RCA-I+ (binding of 16 or 29%, for I or II, respectively), WGA+(8 or 7%) [530] Seal intestine of the suckling animals (Phoca groenlandics)

Alkaline phosphatase 260kDa, SU 67kDa Lectin-Sepharose (sequential ConA+ (desorption with chromatography) 50mM MM), ConA-WGA+, ConA-PSA+, ConA-RCA-I- Carbohydr.(for ConA+-form): 23% (including Man, Gal, GlcN) [531,532]

(3.1.3.1.) Protein inducer of alkaline phosphatase Bovine fetal serum Monomeric factor 64.5 kDa inducing alkaline phosphatase in primary cultures of hen epiphyseal growth plate chondrocytes Cibacron-Blue-agarose—QMA-anion- exchange-Silica (Accell), ConA-Sepharose ConA+ (for purification) [533] (3.1.3.1.) Protein converting alkaline phosphatase form M into forms A and B Human serum Factor 60kDa converting alakline phosphatase M into A and B (without proteinase, lipoprotein lipase or lecithin cholesterol acyltransferase activity) DEAE-Sepharose—ConA-Sepharose — ConA+ (complete binding; UltrogelAcA34 desorption with 50mM MG) [534] 3.1.3.2. Acid phosphatase

Bacteria Mycoplasma faucium IID996,M.fermentans IID812 (human mycoplasmas,cell Mb)

Acid phosphatase 31,2kDa (SDS-PAGE) solubilized with TX100, specific for O-phospho-L-Tyr CM-Sepharose**--ConA-Sepharose — ConA+ (desorption with CM-Sepharose gradient 0-1 M MM; recovery of 33%) [535] Lower plants Aspergillus ficuum NRRL 3135 (SRRC 265)

(fungal CF)

Acid phosphatase 130kDa, SU 68kDa, PAS+ pI 4,0 DEAE-sorbent—SP-Trisacryl—Accel-QMA- Exchanger—SP-Trisacryl** Carbohydr.: 52%; M/M: 34 Gal, 116 Man, 42 GlcN(Ac); Asn-glycans of oligoMan type EndoH+ [536]

Pleurotus ostreatus (edible mushroom fruiting bodies) Acid phosphatase HA—Mb-ultrafiltration(repeated) Activation with endogenic lectin POA POA+ (15%; Gal as inhibitor) [537] Rhodotorula glutinis (yeast CF) Purified acid phosphatase 93kDa (SDS- PAGE),and its deglycosylated form 76kDa LCA-SepharoseCL4B LCA+ (desorption of >95% 0,15M MM; separation from exogenic alpha-mannosidase) [538] EndoH-treated-enzyme—SDS-PAGE—125I- ConA ConA+ (O-linked mannan) [539] Carbohydr.: 40% (80% of total carbohydr. in 11 Asn-glycans, and 20% in 15 Ser/Thr- glycans) [538-540]

Saccharomyces cerevisiae (yeast protoplasts and cell walls) Acid phosphatase forms containing 70, 53, 45, or 38% Man [541]

ConA-precipitation of acid phosphatase forms (43-55% carbohydr.) ConA+ [542]

Acid phosphatase 243kDa, SU 121kDa (or SU 60kDa for deglycosylated enzyme), 50,5% carbohydr. [543]

Stabilization of acid phosphatase by cross-linking of their glycans [23]

Importance of acid phosphatase carbohydr. for enzyme oligomerization (octameriz- ation) and for high level of enzyme activity [544]

Saccharomyces cerevisiae X2180-1A (yeast osmotic shocked cells) Acid phosphatase 140kDa PAS+ Asn-glycans of oligoMan type EndoH+ (appearance of major SU 56kDa, and minor SU 60kDa) [545] S. cerevisiae P28-24C (yeast

34 protoplasts, CF) Acid phosphatase forms 110-180kDa, or 59+57+55kDa from tunicamycin-treated cells ConA-Sepharose ConA+ (complete binding) Asn-glycans of oligoMan type EndoH+ (110-180kDa) EndoH- (59+57+55kDa) [546] S. cerevisiae strain 367 (yeast cell walls, CF) Acid phosphatase constituitive form DEAE-SepharoseCL6B**—SephacrylS300, ConA-precipitation ConA+ Carbohydr.: 55% Man, 2.5% GlcNAc [547]

Higher plants Arachis hypogaea L. (peanut cotyledon plasma Mb) Mb-bound acid phosphatase with SU 46,7+ 50,1 kDa DEAEC Carbohydr.: 50% [548]

Acid phosphatase ConA-DEAEC ConA+ (complete binding; desorption with 0,1M Man) [467] Brassica nigra (black mustard leaf petiole cell-suspension culture) Monomeric acid phosphatase 60kDa, pI PAS+ 4,5 S-Sepharose(*)**— Chelating-Sepharose — ConA-Sepharose —Superose12* —Phenyl- ConA+ (desorption with Superose* gradient 0-0,5M MM; 65%, 1,3-f) [549] Canavalia ensiformis L. (jack bean) Intracellular acid phosphatase ConA-Sepharose ConA+ (desorption with 0,1M MM) [550] Cucurbita ficifolia (roots of seedlings) Crude acid phosphatase Inhibition with endogenic lectin CFL CFL+ (30-40%) [551]

Acid phosphatase forms Ba (associated with CFL), and Bb ConA-Sepharose ConA+ (desorption of Ba with 0,15mM MM followed by 0,3M MM for Bb) [551] Dactylis glomerata (seeds) Acid phosphatase of extract Inhibition with lectin ConA+ (50%) [552]

Glycine max L.(soybean, germinating seed axes) Monomeric acid phosphatase 50kDa CM-Sephadex —ConA-Sepharose** —HA ConA+ (desorption with gradient 0-0,5M Man; 76%, 10-f) [553]

Glycine max L. (seed germinating cotyledons) Acid phosphatase SP-Toyopearl —ConA-Sepharose4B** —HA ConA+ (desorption with gradient 0-0,5M Man; 45%, 10-f) [553] Glycine max L. Merr. (soybean root- derived nonphotosynthetic cell line SB1, cell walls)

Ionically bound cell wall purple acid phosphatase 130kDa,SU 58kDa,pI 5.0, solubilized with 0.5M CaCl2 CMC** —ConA-Sepharose —BiogelA1.5m ConA+ (desorption with 1M MM and 0,5M Glc; 39%, 1,6-f) [554] Glycine max L. (cells in culture, cell walls) Homogenic cell wall acid phosphatase (SDS-PAGE) Dot-Blot —Lectin ConA3+, GNA3+, TPA+, SBA+ [431] Gossypium hirsutum var.Bikaneri Nerma Acid phosphatase 200kDa, SU 55kDa CM-Sephadex —ConA-agarose** ConA+ (desorption with 0,5M MM; 15%, 39-f) [555] Helianthus annuus L. (sunflower seeds) Acid phosphatase 103kDa, SU 56+52kDa ConA-Sepharose** —Affigel-Blue — ConA+ (desorption with SephacrylS200 0,4M Man; 61%, 39-f) [556] Hordeum sativum (barley roots) Acid phosphatase forms A, Ba1, Ba2, Bb1, and Bb2 ConA-Sepharose —SP-Sephadex —QAE- ConA- (A), ConA+ (desorption Sephadex of Ba with 15mM MM followed by 0,5M MM for Bb) [557,558] Precipitation with lectins in agarose ConA3+, LCA+, WGA-, SBA- [557] Acid phosphatase forms B1 and B2 ConA-Sepharose —SephadexG200(or BiogelP100)—Phenyl-Sepharose ConA+ (desorption with 0,5M MM) [558,559] LAE in agarose ConA+ (differences between B1 and B2) [559] Hordeum vulgare var.Beca (barley germinating seeds) Acid phosphatase 57kDa (SDS-PAGE) PAS+ DEAEC Carbohydr.(mkg/mg protein): 490 (total hexoses); 455 (Gal), 6 (GlcN), 5 (Sia) [560]

Ipomoea batatus (sweet potato tubers)

Mn(III)-acid-phosphatase 110kDa, SU 55

36 kDa, pI 5,18 (violet enzyme) DEAE-Sephadex—DEAE-Sephadex —ConA- ConA+ (desorption with Sepharose 0,1M MM) [561]

Fe-acid-phosphatase 113kDa, SU 53kDa, pI 5.14 (purple enzyme) DEAEC—ConA-Sepharose**—Sephacryl- ConA+ (desorption with S300 gradient 0-0,3M Man; 88%, 12-f) [562] Lupinus angustifolius (blue lupin seeds, stems, and cotyledons) Grass-Ba1-like acid phosphatase ConA-Sepharose —BiogelP100 —SP- ConA+ (desorption with Sephadex 0,3M MM)

Precipitation with ConA in solution ConA+ (active precipitate)

Enzyme-Lectin complex in agarose ConA+, LCA+, SBA-, WGA- Asn-glycans of oligoMan type EndoH+ [563]

Lycopersicon esculentum Mill. cv.VFNT (tomato cell-suspension culture) Acid phosphatase forms 1 (47kDa), PAS+ (form 1) and 2 (SU 48+56kDa) DEAEC—HA—ConA-Sepharose ConA- (for 2), ConA+ (for 1; desorption with 50mM MG) [564] Oryza sativa L. Japonica cv.Koshihikari (rice grain aleurone particles) Violet acid phosphatase 72kDa (gel filtartion), or 68kDa (SDS-PAGE) PAS+ DEAE-Sephadex—ConA-Sepharose — ConA+ (desorption with 0,25 DEAEC**—SephadexG200 M MG; 132%, 7-f) [565,566] Carbohydr.: Glc-beta- Cellulosine+, EndoH- [565] Oryza sativa L. (rice bran) Acid phosphatase forms 1, and 2 (both with ) DEAEC—Phosphocellulose**—ConA- ConA+ (desorption with Sepharose—Sepharose 0,2M MG; 30%, 6,0-f; desorption of 1 with 50mM MG followed by 100mM MG for 2) Acid phosphatase forms 3,and 4 (both without phytase) DEAEC—CM-Sephadex—CM-Sephadex— ConA-Sepharose**—Sephadex ConA+ (desorption with borate; 35%, 13-f) [567] Phaseolus vulgaris L.(red kidney bean) Dimeric Fe(III)-Zn(II)-purple-acid- phosphatase 130kDa SephacrylS200—ConA-Sepharose ConA+ (desorption with 0,25 M MM; 1.8-f; irreversible binding without gel- filtration step) [568]

5 Asn-glycans of complex type with PNGaseA+, PNGaseF- Xyl (including 4 glycans with core Fuc-alpha-3-) [569]

Poa pratensis L. (grass seeds) Acid phosphatase forms I and II ConA-Sepharose ConA+ [558,570] LAE in agarose ConA2+, LCA+, WGA-, SBA- [570,571] Activation with lectins ConA2+, LCA+, WGA- [552,570] ConA-Sepharose(ConA-)—BiogelP100— ConA-Sepharose—Phenyl-Sepharose ConA+ (binding of >80%; desorption with 0,5M MM) [571] Carbohydr.: 40 or 20%, for I or II, alpha-L-Fucosidase+, respectively alpha-Glucosidase+, beta-Galactosidase+ [571] Poa pratensis var.Skrzeszowicka (grass caryopses, stored from 1 to 10 years at 20oC)

Acid phosphatase Lectin-Sepharose ConA+ (partial binding; increased binding for 10-year caryopses), WGA-, Activation with ConA ConA+ (decreased binding upon storage) [572] Poa pratensis (grass roots of 7-days seedlings) Acid phosphatase forms Ba1,Ba2,Bb1, and Bb2 ConA-Sepharose—SP-Sephadex(or QAE- ConA+ (desorption of Ba Sephadex) with 15mM MM followed by 0,5M MM for Bb) [557] Raphanus sativus L. cv.Taibyosobutori (Japanese radish seedlings) Acid phosphatase 63kDa (TSKgel- G3000SW*) or 58kDa (SDS-PAGE), pI 6,9 Q-Sepharose*—ConA-Sepharose—IEF**— ConA+ (desorption with TSKgelG3000SW* 0,5M MM; 70%, 2,7-f) [573] Secale cereale (rye seed germs) Acid phosphatase forms 90, 60, 35kDa, SU (30+60kDa), pI 6-6,4, 5,4-5,6 SP-Sephadex—ConA-Sepharose—Biogel- ConA+ (desorption with P150 0,25M MG) Activation with lectins STA+ (55%), ConA+ (46%), LCA+ (30%), SBA+(19%), WGA+ (15%) [552,574] Solanum tuberosum L. (potato tubers) Acid phosphatase of extract Activation with lectins STA+ (30%), ConA- [552]

Acid phosphatase 69kDa DEAEC—BiogelP150—ConA-Sepharose** ConA+ (desorption with 0,25M MM; 90%, 5-f) [575]

38

Activation with lectins STA+ (39%), ConA- [576] Carbohydr.: 16.6% (5.6% Man, 3.4% Rha, 2.5% Glc, 1.5% Gal, 3.6% GlcN) [575]

TriticoSecale (X. TriticoSecale Wittmack, winter triticale, plump or shriveled seeds) Acid phosphatase 45,7 kDa (SDS-PAGE), pI 5,9 DEAE-Sephacel—Chromatofoc— ConA-Sepharose4B—BiogelP60** ConA+ (desorption with 0,2M MG; 93%, 1,9-f, for plump seeds; 88%, 1,8-f, for shriveled seeds) Carbohydr.: 12% [577]

Triticum vulgare (wheat grains, germs, leaves, roots) Acid phosphatase of the grain extract WGA-Sepharose WGA+ (desorption with 0,2M GlcNAc) [578] Monomeric acid phosphatase 58kDa of wheat germs SephadexG75—SP-Sephadex—DEAEC— SP-Sephadex, ConA-Sepharose ConA- Carbohydr.: <1% [579]

Acid phosphatase forms B1 and B2 of leaves LIE in agarose ConA+ (releasing from precipitates with 0,12 or 0,25M MM, for B1 or B2, respectively) [558] Acid phosphatase forms Ba1,Ba2,Bb1, Bb2 of 7-days seedling roots ConA-Sepharose—SP-Sephadex(or QAE- ConA+ (desorption of Ba Sephadex) with 15mM MM followed by 0,5M MM for Bb) LIE in agarose ConA3+ (all isozymes), LCA+ (traces of precipitates of all isozymes), WGA-, SBA- [557] Protozoans Entamoeba histolytica NIH-200 (amoeba, CF) Acid phosphatase forms I, II, III DEAEC Modulation of activity with ConA ConA+ (inhibition of 38%, for I; activation of 102 or 108%, for II or III, respectively) [579] Leishmania braziliensis, L. mexicana (leishmanial promastigote Mb) Citrate-sensitive acid phosphatase form Sepharose4B—DEAE-Sephacel—ConA- ConA+(desorption with Sepharose**—IEF 50mM MM; 72%, 14-f)

Citrate-resistant acid phosphatase form solubilized with TX100 Sepharose4B—DEAE-Sephacel**—ConA- ConA+ (desorption with Sepharose 50mM MM; 57%, 5,7-f) [580]

Leishmania donovani strain Sudan 1 (promastigote Mb) L(+)-tartrate-resistant acid phosphatase 128kDa,SU 65+68kDa, pI 4.1,solubilized with Na-cholate

SephadexG75—ConA-Sepharose—QAE- ConA+ Sephadex—Phenyl-Sepharose— SephadexG100 [581]

SephadexG75—QAE-Sephadex—ConA- ConA+ (desorption with Sepharose**—SephadexG150(Sephadex- 0,2M MM; 78%, 8,3-f) G100) [582]

L(+)-tartrate-sensitive acid phosphatase forms 132kDa, pI 5,4, and 108kDa, pI 7,1 SephadexG75—QAE-Sephadex—ConA- ConA+ (desorption with Sepharose 0,2M MM) [582]

Leishmania donovani strain 1S (promastigotes, CF) Tartrate-sensitive acid phosphatase 134kDa, pI 4,5 LCA-Sepharose4B LCA+ (desorption with 0,3M MM; 72%, 16-f) Carbohydr.: 37% [583]

Leishmania donovani strain Sudanese (promastigotes, CF) Acid phosphatase forms 149 and 97kDa (SDS-PAGE) RCA-I-agarose—LCA-Sepharose4B RCA-I-LCA+ (desorption of 70% with MM) [584] L. donovani (promastigotes, CF) Acid phosphatase forms 130 and 110kDa (SDS-PAGE) Sepharose4B—Lectin-agarose ConA+, LCA+, RCA-I+, PNA+ (binding >90% to each sorbent; desorption with 0,5M sugar) [585] Leishmania major, L. mexicana (promastigote lysosomal Mb) Acid phosphatase forms 72 and 70kDa (SDS-PAGE) solubilized with OG ConA-Sepharose—mAb(AP4)-Sepharose ConA+(desorption with 0,5M MM) Carbohydr.: 29-30% Asn-glycans: 20-22% in 70-72kDa EndoF+ ([72+70]—50kDa) [586]

40

Leishmania tropica LRC-L36 (promastigotes, CF) Tartrate-sensitive acid phosphatase 200kDa mAb(T15)-Sepharose4B—LCA-B-agarose LCA-B+ (desorption with (Bio-Makor, Israel) 0,2M Man; 50%, 2,0-f; separation from unbound phosphoglycans) [587] Invertebrates Apis mellifera (honeybee, liophylezed venom)

Acid phosphatase ConA-Sepharose4B ConA+ (complete binding; desorption with 0,5M MM) [387] Monomeric acid phosphatase 62kDa, pI 5,1, forms I and II SephadexG75—DEAEC(I+II)—Sephadex- G200, ConA-Sepharose ConA+ (two fractions), ConA- (one fraction) Carbohydr.: 2,8 or 6,4%, for I or II, respectively) [588]

Acid phosphatase forms 96 and 45kDa PAS+ (SDS-PAGE) SephadexG100**—CM-Sephadex [589]

Eimeria tenella (hen parasitical helminth, sporulated oocysts) Acid phosphatase 44kDa (SDS-PAGE), forms I and II DEAEC(I+II)—SephadexG200—ConA- ConA- (for both forms) Sepharose Carbohydr.: Sia [590]

Paracentrotus lividus (sea urchin eggs) Acid phosphatase particulate form 600kDa (Sepharose 4B) UltrogelAcA34—DEAEC—HA— PAGE**, ConA-Sepharose4B ConA+ (complete binding) ConA- (in the presence of 0,2M MM) [591] Spaerechinus granularis (mediterranean sea urchin eggs) Acid phosphatase particulate 700kDa (Sepharose4B) UltrogelAcA34—DEAEC—UltrogelAcA34 —DEAEC—HA—PAGE**, ConA-Sepharose4B ConA+ (complete binding without desorption by 0,2M MM or 0,5M borate), ConA- (in the presence of 0,2M MM) [591] Fishes Catfish liver Acid phosphatase forms I and II ConA-Sepharose4B—Chromatofoc ConA+ (for both forms;

desorption with 0,25M MG) Carbohydr.: 3,6 or 21%, for I or II, respectively [592]

Scomber japonicus (common mackerel, dark muscle) Acid phosphatase hydrolizing pNP- monophosphate ConA-Sepharose ConA- (majority of activity), ConA+ (desorption of minor form with 50mM MM) [593] 5'-Inosinate-monophosphatase (acid IMPase) 90kDa (SephadexG100) ConA-Sepharose**—DEAEC ConA+ (washing with 50mM MM; desorption with 0.5M MM; 44%, 157-f; separation from unbound acid phosphatase) [593] Amphibium Rana esculenta (frog liver) Acid phosphatase forms 140 and 38kDa (isoforms I-IV of around 38kDa) ConA-Sepharose—BiogelP200 ConA+ (around 38kDa) [594] Monomeric acid phosphatase III (35,1 kDa, pI 5,87) and IV (35,5 kDa, pI 6,24) ConA-Sepharose—BiogelP200— ConA+ (all low mol.weght Chromatofoc—SephadexG75 forms)

Activation with ConA ConA+ (40% for III, 28% for IV) [595] Acid phosphatase forms I-IV Chromatofoc—Dot-Blot—Lectin ConA2+ (I, II), ConA+ (III, IV), WGA2+ (III), WGA+ (I, II, IV), LCA+ (IV), PNA+ (low affinity of IV), PNA2+ (Sialidase-treated forms), WGA- (decreased affinity of Sialidase- treated forms) Carbohydr.: Sia-alpha-Gal- Sialidase+ [596]

Mammals Human liver Acid phosphatase 90-93kDa,SU 50-52kDa CMC**—DEAEC—ConA-Sepharose4B— ConA+ (desorption with SephacrylS200 gradient 0-0,2M Man; 92%, 3,1-f) Carbohydr.: 4% (2.9% Man,1.1% GlcNAc) [597]

Human osteoclastoma Acid phosphatase 30kDa (Superose12*) CM-Sepharose**—Phenyl-Sepharose— ConA-Sepharose—Superose12*—MonoS*— ConA+ (desorption with Superose12* 0,4M MM; 59%, 5,5-f)

42

[598] Human placental fibroblast lysosomes Tartrate-sensitive acid phosphatase 90kDa, SU 48kDa DEAEC—CMC—ConA-Sepharose**—IEF ConA+(desorption with —SephacrylS200—HA—SephacrylS200 0,5M MG; 74%, 25-f) [599,600] At least 1 Asn-glycan of oligoMan type [600]

Tartrate-sensitive acid phosphatase forms A1(98kDa), A2(93kDa), B(93kDa), and tartrate-resistant form C (>200kDa) ConA-Sepharose—L-Tartrate- ConA- (41%, for C), ConA+ Sepharose—DEAEC (partial separation of A1, A2, B using gradient 0-50mM MM; 40%, 40-f, for A1; 41%, 32-f, for A2; 41%, 28-f, for B) [601] Human prostatic adenoma Acid phosphatase of extract ConA-Sepharose ConA+ (washing with 2mM MM; desorption with gradient 2-50mM MM; 67%, 3,8-f) [602,603] ConA-Sepharose (for biocatalyst) ConA+ (immobilization of 80% active enzyme) [604] ConA immobilized in microplate ConA+ wells [605]

CM-Sephadex**—ConA-Sepharose— ConA+ (desorption with SephacrylS200 0,5M MG; 102%, 5,0-f; separation from haptoglobin SU) [606] Acid phosphatase 100kDa, SU 48kDa, sialoforms pI 4,8 and 4,5 ConA-Sepharose**—DEAEC—SephadexG100 ConA+ (desorption with —SephadexG100 gradient 0,1-0,5M MM; 89%, 5,5-f) [607] Acid phosphatase groups I (11-13 forms, pI 3,8-4,8), Ia (SU 46+50kDa, 9 forms pI 3,8-4,7), Ib (SU 76kDa, 2-4 forms pI 4,5-4,8), II (SU 46+50+76kDa, 4-8 forms pI 5,0-5,5), III (SU 86+76kDa, 5 forms pI 5,6-5,9) UltrogelAcA44—IEF—SDS-PAGE—ConA- ConA+ (for cytosolic 50 peroxidase and 46kDa)

Carbohydr.: Sia C.perfringens or Vibrio cholerae Sialidase+ (group I, and few forms of II) [603,608] Acid phosphatase 100kDa Carbohydr.: 12.7% (6.4% hexoses, 4.7% hexosamines, 1.6% NeuNAc) [609]

Human seminal fluid

Acid phosphatase 50kDa (SephadexG75) or 45kDa (SDS-PAGE) ConA-Sepharose4B—DEAEC—Sephadex- ConA+ (93%, 6,8-f) G150—SephadexG75—SephadexG150 Carbohydr.: 15% (8.4% hexoses, 5.3% hexosamines, 1.3% NeuNAc) [609]

Acid phosphatase group I (SU 46+50kDa, 9 forms pI 3,8-4,7) UltrogelAcA44—IEF—SDS-PAGE—ConA- ConA2+ (50kDa), ConA+ peroxidase (46kDa) Carbohydr.: Sia Sialidase+ [608]

Acid phosphatase forms A and B SephadexG200(A+B), ConG-agarose ConG+ (desorption with 20 mM MM; 54-f by the use of gradient 0-0,1M MM) [610] Activation with seed extract containing lectins Phaseolus coccineus+ (50%), Vigna Catiang Endl.var. Sinensis+ (35%), Cytisus scorparius Link+ (31%), Wistaris branchbotrys Sieb et Zucc+ (15%) [610] Inhibition with seed extract Canavalia gladiata+ (70%) containing ConG [610]

Human serum of patients with bile duct ligation or hepatocellular injury Acid phosphatase Inhibition with ConA ConA+ (up to 30%) [611]

Human spleen from patients with Gaucher's disease Tartrate-sensitive acid phosphatase SP-Sephadex—QAE-Sephadex—DEAEC—HA —ConA-Sepharose—SephadexG200 ConA+ (desorption with 0,2M MM; 97%, 4-f) Carbohydr.: Sia Sialidase+ [612]

Kurloff cell bodies Urea-soluble acid phosphatase 30-35kDa (SDS-PAGE) SDS-PAGE—Blot—Lectin-digoxigenin SNA+, RCA-I+ (Sialidase- (or lectin-biotin) treated enzyme), WGA+, PHA-E+, ConA+ Sialylated Asn-glycans of complex Sialidase+ , PNGaseF+, type O-Glycanase- [537]

Porcine endometrium cells in culture Uteroferrin (Fe-containing purple acid phosphatase) forms 35 and 37kDa Mixture of Gp Lectin-Sepharose ConA+, LCA+, WGA+ (separation of Gp)

44

Asn-glycans of oligoMan type (35kDa); EndoH+ (35 and 37kDa) two glycans of oligoMan type and complex type (37kDa) [613]

Rabbit kidney cortex Tartrate-sensitive basic major form 54kDa (SDS-PAGE) or 101kDa (ultracentrifugion), and acid minor form SP-Sephadex—DEAEC**— ConA-Sepharose ConA+ (desorption with 0,6M MM; 89%, 3,5-f, for basic form; 69%, 1,9-f, for acidic form) [614] Rat bone osteoclasts Tartrate-resistant acid phosphatase, or recombinant purple acid phosphatase Lectin-sorbents Lectin+ (purification) [614a]

Rat developing bone Monomeric purple acid phosphatase with two reduced polypeptides 32+16kDa (SDS-PAGE, Me-treated protein) CMC—SephadexG75—SephacrylS200— ConA-Sepharose ConA+ (desorption by 0,5 M MM with 30% ethylene- glycol; 75%, 6,2-f) [615] Rat liver lysosomes Soluble acid phosphatase forms I, II, and III ConA-Sepharose**—DEAEC—CRA- ConA+ (for all forms; Sepharose desorption with 0,13M MM; 60%, 24-f), CRA- (38%), CRA+ (separation of I-III using gradient 0-35nM sialodisaccharide from the sheep submaxillary mucin; desorption with 4-8, 14- 17, or 23-25nM sugar,for I, II, or III, respectively) Sia (nM/mg protein): 30,56,or 89, for I, II, or III, respectively [616]

Tartrate-resistant and tartrate- sensitive acid phosphatase forms A and B, respectively RCA-I-Sepharose4B RCA-I+ (desorption with 0,25M Lac; 5,9-f, for A; 9.4-f, for B) [617] Rat spleen Monomeric Fe-containing purple acid phosphatase 33kDa associatedwith phosphoprotein phosphatase Blue-Sepharose—ConA-Sepharose**— ConA+ (desorption with SephadexG100—CM-Sephadex 0,5M MG; 76%, 28-f) [618]

3.1.3.4. Phosphatidate phosphatase

Rat liver plasma Mb Phosphatidate phosphohydrolase forms 51kDa, pI 9, and 53kDa, pI<4, solubilized with TX100 For 51kDa: HA—Q-Sepharose*—S- Sepharose*—MonoQ*—WGA-Sepharose WGA+ (desorption of 20-40% —HA with 0,5M GlcNAc) For 53kDa: HA—Q-Sepharose*—Phospho- agarose—MonoQ* Carbohydr.: Sia (2 kDa in 53 kDa), Sialidase+ (53 —51kDa), Sialylated Asn-glycans of complex type (23kDa N-GlycanaseF+ ([51-53]—28kDa) in 53 kDa, 21 kDa in 51 kDa) [619]

3.1.3.5. 5'-Nucleotidase

Lower plant Dictyostelium discoideum (slime mold vegetative cell Mb) 5'-Nucleotidase associated with alkaline phosphatase ConA-Sepharose4B**—DEAE-Sephacel ConA+ (binding of 98%; desorption with 0,2M MM; 79%, 2,2-f) [456,457] Higher plants Arachis hypogaea L.(peanut cotyledon plasma Mb, Golgi apparatus) Plasma Mb-bound 5'-nucleotidase 55kDa (SDS-PAGE) solubilized with OG DEAEC Carbohydr.: 42,7% [620]

5'-Nucleotidase 53,7 kDa (SDS-PAGE) from Golgi apparatus, solubilized with OG DEAEC Carbohydr.: 38,5% [621]

Glycine max L. (soybean root nodules) 5'-Nucleotidase 70kDa forms I (pI 8,5), II (pI 8,2), III, and IV CM-Sephadex**—ConA-Sepharose— ConA+ (all forms; Fractogel—Chromatofoc desorption by 0,3M MM with 20% ethyleneglycol; 53%, 4,8-f) [622] Fishes Gadus macrocephalus (cod muscle) 5'-Nucleotidase 270kDa, SU 67kDa ConA-Sepharose—5'AMP-Sepharose** ConA+(desorption with 0,1M MM; 96%, 17-f) [623] Torpedo marmorata (electric ray, electric organ) 5'-Nucleotidase 131kDa, SU 62kDa ConA- MP-Sepharose** ConA+ (desorption with 0,4M MM; 57%, 3,6-f)

SDS-PAGE—Blot—ConA-peroxidase ConA+ (62kDa)

46

Inhibition with lectins ConA+ (84%), LCA+ (19%), LTA-, PNA-, WGA-, VVA- [624] Birds Hen gizzard muscle Mb Mb-bound 5'-nucleotidase forms 310 and 152kDa, SU 79kDa AMP-Sepharose**—LCA-Sepharose LCA+ (desorption with 0,2 —DEAEC M MM; 89%, 3,3-f) [625,626] SDS-PAGE—Blot—ConA-peroxidase ConA+ (79kDa)

+ Inhibition with ConA ConA (Ki=0.076 mkM) [625] Hydrophilic 5'-nucleotidase solubilized with phospholipases C and D Inhibition with ConA (20mkg/ml) ConA+ (90 or 20%, for phospholipase-treated, or native enzyme, respectively) [627] Mammals Bovine caudate nuclear synaptic plasma Mb Plasma Mb-bound 5'-Nucleotidase (I) or enzyme form solubilized with zwitterionic detergent sulfobetaine-14 (II) For II: Lectin-UltrogelAcA44 WGA+ (desorption with 68 mM GlcNAc using gradient 0-0,2M sugar), PNA-, RCA-I- Sialidase-treated II (III) Lectin-UltrogelAcA44 WGA+ (desorption with 34 mM GlcNAc), PNA+or RCA-I+ (desorption with 0,2M Gal)

Inhibition of III with lectins LCA+ (80%), WGA+(56%), PNA+ (51%), UEA-I- beta-Galactosidase-treated III (IV) Lectin-UltrogelAcA44 WGA+ (desorption with 52mM GlcNAc), PNA-, RCA-I-

Inhibition of IV with lectins LCA+ (74%), WGA+(63%), PNA-, UEA-I- beta-N-Acetylglucosaminidase-treated IV (V) WGA-UltrogelAcA44 WGA+ (desorption with 28mM GlcNAc) Inhibition of V with lectins LCA+ (80%), WGA+ (24%), PNA-, UEA-I- alpha-Mannosidase-treated V (VI) WGA-UltrogelAcA44 WGA+ (desorption with 48mM GlcNAc) Inhibition of VI with lectins LCA+ (82%),

WGA+ (53%), PNA-, UEA-I- III treated with EndoH, EndoD (VII) UEA-I-UltrogelAcA44 UEA-I+ (desorption with 0,2M alpha-L-Fuc) Inhibition of VII with lectins LCA+ (80%), WGA+ (77%), PNA-, UEA-I- II treated with EndoD followed by alpha-L-fucosidase (VIII) UEA-I-UltrogelAcA44 UEA-I-

Inhibition of VIII with lectins LCA+ (75%), WGA+ (72%), PNA-, UEA-I-

Inhibition of I and II with lectins ConA+ (85 or 80%, for I or II, respectively), LCA+ (80 or 75%), LPA+(75 or 65%), WGA+(80 or 75%), PNA-, RCA-I-, UEA-I- [628] Bovine hepatocyte plasma Mb Mb-bound 5'-nucleotidase Inhibition with lectins ConA+ (90%), LCA+ (25%), WGA+ (50%) [414] Bovine liver Solubilized of Mb-bound 5'-nucleotidase (SU 70kDa) ConA-Ultrogel** —AMP-Sepharose — ConA+ (84%, 10-f) UltrogelAcA54 [629]

Solubilized (I) and Mb-bound (II) enzyme forms Inhibition with lectins For I: LCA2+, ConA+, WGA+ For II: ConA2+, LCA+, WGA+ [629] Cytosolic heterodimeric 5'-nucleotidase 130kDa ConA-Sepharose —AMP-Sepharose** ConA+ (washing with 10mM MG; desorption with 0,3M MM; 55%, 33-f; 10-f if without washing by 10mM MM) Inhibition with lectins ConA+ (85%), LCA+ (30%), WGA- [630] Bovine lymph node lymphocyte plasma Mb Dimeric 5'-nucleotidase (5.5S) Inhibition with lectins ConA+ (90 or 50%, for intact or alpha-mannosidase- treated Mb, respectively), WGA+ (40%,for alpha-mannosidase-treated Mb), PNA- or WGA- (for intact Mb), LCA+ (85%, for intact Mb), UEA-I-

48

Carbohydr.: Man-alpha- alpha-Mannosidase+ [414]

Bovine milk fat globule Mb 5'-Nucleotidase solubilized with DOC Inhibition with ConA ConA+ (non-competed cooperative binding) [631] Bovine retinal rod Mb Monomeric 5'-nucleotidase 75kDa SephacrylS300 —AMP-Sepharose**

Mb-bound enzyme Inhibition with lectins ConA+ (non-competed, ki=3 mkg/ml), WGA+ (non- competed, ki=58 mkg/ml), RCA-I+ (competed, ki=1,2 mkg/ml), (ConA+WGA)+, (ConA+RCA-I)+ [632] Bovine seminal plasma, spermatozoides 5'-Nucleotidase 160kDa (BiogelA1.5m), SU 72kDa DEAE-Sephadex —ConA-Sepharose4B — ConA+ (desorption with ADP-agarose** 0,4M MM; 73%, 4,4-f) [633] Mb-bound (I) and solubilized (II) with detergent enzyme forms + Inhibition with lectins ConA (ki=14 or 270nM, for I or II, respectively) [634] ConA+ or LCA+ (90%) [635] Bovine thyroidal plasma Mb 5'-Nucleotidase 150kDa, SU 75kDa, solubilized with Na-DOC LCA-Sepharose4B** —5'AMP- LCA+ (desorption with Sepharose4B 0,13M MM; 81%, 9,5-f; separation from unbound alkaline phosphatase), LCA- (up to 25%) [474] Mb-bound (I) and purified solubilized (II) enzyme forms Inhibition with lectins ConA+ (70 or 55%, for II or I, respectively), LCA+ (49%, for I), WGA+ (4%, for I), sConA+ (3.5%, for I) [474] Feline Schwann cell plasma Mb of degenerated nerve Mb-bound 5'-nucleotidase Inhibition with ConA ConA+ (up to 80%) [636]

Guinea pig skeletal muscle plasma Mb Mb-bound (I) and solubilized (II) 5'-nucleotidase forms + Inhibition with ConA ConA (ki=50 or 160nM,

for I or II,respectively) [634] Human colon adenocarcinoma cells BCS- TC2 Plasma Mb-bound 5'-nucleotidase Inhibition with ConA ConA+ (60 or 75%, for monolayer or suspension cells, respectively) [637] Human gingival hyperplasia cells FB-HG Plasma Mb-bound 5'-nucleotidase Inhibition with ConA ConA+ (90%, in cell suspension) [637] Human glioblastoma Rugli cell plasma Mb Mb-bound 5'-nucleotidase Inhibition with ConA ConA+ (85 or 95%, in monolayer or suspension cells, respectively) [637] Human pancreatic adenocarcinoma cell plasma Mb Mb-bound 5'-nucleotidase solubilized with Bacillus thuringiensis phospho- lipaseC or mammalian phospholipaseD Inhibition with ConA ConA+ (increased effect for phospholipase-treated enzyme) [627] Human placenta Microsomal 5'-nucleotidase solubilized with TX100 and Na-DOC DEAEC** —SephadexG100 —SephadexG200 —DEAE-BiogelA Inhibition with lectins ConA2+ (80%), LCA+ SDS-PAGE Dansyl-hydrazine+ [638]

Plasma-Mb-bound 5'-nucleotidase solubilized with phosphatidyl-inositol- specific phospholipaseC, SU 73kDa ConA-Sepharose**—AMP-Sepharose ConA+ (desorption with 0,3M MM; 68%, 7,2-f) [639] Cytoplasmic 5'-nucleotidase 143kDa, SU 76kDa 5'AMP-Sepharose-4B**—BiogelP300— ConA-Sepharose ConA+ (desorption with 10mM MM; 100%, 15-f) [640] Human seminal plasma 5'-Nucleotidase with SU 71kDa (SDS-PAGE) SephacrylS300—ConA-Sepharose—AMP- ConA+ (washing with 0,1M agarose** MG; desorption with 0,3M MM; 63%, 2,7-f; separation from acid phosphatase) [641] Human serum 5'-Nucleotidase Inhibition with ConA ConA+ (80-90%) [611]

Murine ascites Erlich-Lettre tumor cells containing or noncontaining glycogen

50

(I- or II-type cells, respectively) Plasma-Mb-bound 5'-nucleotidase from I or II Inhibition with ConA ConA+ (95 or 60%, for I or II, respectively) [642,643] Murine ascites plasmacytoma MOPC 173 cell plasma Mb Mb-bound 5'-nucleotidase forms I and II separated with zonal rotor centrifugion Inhibition with ConA ConA+(52 or 60%,for I or II,respectively) [644] Murine ascites tumor cells ELT Bonn cultivated in lipid-depleted medium Plasma-Mb-bound 5'-nucleotidase forms I and III SephacrylS1000 Inhibition with ConA ConA+ (16 or 41%, for I or III, respectively) [645] Murine thymocyte plasma Mb Mb-bound 5'-nucleotidase Inhibition with ConA ConA+ [646]

Porcine mesenteric lymph node lymphocyte plasma Mb Mb-bound 5'-nucleotidase solubilized with dodecyltrimethylammonium-bromide (DTAB) LCA-Sepharose 4B LCA+ (desorption with 0,2M MM; >7-f) [647] Purified enzyme before (I) or after (II) its reassembly into lipid vesicles Inhibition with lectins ConA+ (93 or 77%, for I or II, respectively; KHill = 2.1, for II) [647,648] Mb-bound enzyme solubilized with Na-DOC Lectin-Sepharose4B LCA- (15%), LCA+(desorption with 0,1M MG; 85%, 6,5-f), ConA+ [649]

LCA-Sepharose4B—5'AMP-Sepharose4B**, LCA+ (95%, 6-f), ConA-Sepharose4B ConA+ (recovery 20%) [650]

Rabbit platelet plasma Mb Mb-bound 5'-nucleotidase (GP) Stimulation with ConA ConA+ (2-f or 85%, for intact cells or in the presence of coformycin, respectively) [651] Rat brain and skeletal muscle Partially purified 5'-nucleotidase Inhibition with ConA ConA+ (92%) [652]

Rat forebrain Synaptic plasma-Mb-bound 5'-nucleotidase

Inhibition with lectins ConA+ (70%), WGA+(15%), LPA+ (3%) Cytosol enzyme forms 268kDa, SU 131 kDa, and 286kDa, SU 72kDa Ab-Sepharose4B**—LCA-Sepharose4B LCA+ (desorption with 0,25M MM; 25%, 3,0-f) Inhibition of cytosol enzyme with lectins ConA+ (68%), WGA+ (27%), LPA- Carbohydr.: Glc (decreased activity Glucosidase+ of glucosidase-treated cytosol or plasma-Mb-bound form) [653]

Rat glioblastoma Rugli cell plasma Mb

Mb-bound ecto-5'-nucleotidase forms 135 and 268kDa, SU 74kDa (SDS-PAGE) solubilized with TX100 and CHAPS ConA-Sepharose**—AMP-Sepharose ConA+ (desorption with 0,4M MM; 37%, 34-f)

Inhibition with lectin (50nM) ConA+ (49%), LCA+(38%), PWM-, WGA- [654] Rat heart Mb 5'-Nucleotidase of extract Inhibition with ConA ConA+ (95%) [652] Sarcolemma-bound enzyme forms HL (right-side-out orientation) and S (inside-out orientation) Inhibition with ConA ConA+ (95%, for both forms; KHill = 1,1 or 2,2, for HL or S, respectively [655] 5'-Nucleotidase with SU 74kDa CM-Sephadex—ConA-Sepharose—DEAEC ConA+ (desorption with —ADP-agarose 0,3M MM; 123%, 13-f) [656]

Rat kidney Mb 5'-Nucleotidase of extract ConA-Sepharose ConA+ (binding of 98%; desorption of 103% with 0,4M MM) [657]

Inhibition with ConA ConA+ (96%) [652] Enzyme with SU 69kDa ConA-Sepharose**—ADP-agarose ConA+ (desorption with 0,4M MM; 94%, 17-f) [658] Rat liver plasma Mb, lysosomes Mb-bound 5'-nucleotidase Inhibition with lectins ConA+, RCA-I+, WGA+ [652,659] ConA-Sepharose ConA+(separation from microsomal and mitochon- drial Mb-bound proteins) [660,661] SephadexG100—DEAE-Biogel—HA Inhibition of purified enzyme with lectins ConA+ (94%), sConA+ (8%), RCA-I+ (66%), WGA+ (31%),

52

LTA-, PHA-L-, SBA- [659] Mb-bound 5'-nucleotidase with SU 70+38 kDa (70kDa as catalytic SU) Ab-sorbent—SDS-PAGE—ConA-125I ConA+ (70kDa) [662]

Lysosomal 5'-nucleotidase 140kDa, SU 72kDa, forms pI 4,4-6,0 DEAEC—ConA-Sepharose—Ab-Sepharose- ConA+ 4B—5'AMP-Sepharose4B [663] Carbohydr.: Sia (5.5% in 72 kDa, or 4kDa in Sialidase+ (72—68kDa; 72kDa) pI [4,4-6,0]—[7,4+7,7]) [664] Rat mammary adenocarcinoma cells 13762 MAT-C1 Plasma-Mb-bound (I) and cytoskeleton associated (II) 5'-nucleotidase forms Inhibition with ConA ConA+ (non-cooperative or cooperative, for I or II, respectively) [665] Rat mammary gland microsomal Mb Mb-bound 5'-nucleotidase light (I) and heavy (II) forms separated with sucrose gradient centrifugion + Inhibition with ConA ConA (KHill=1,4 or 1,9- 2,4, for I or II, respectively) [666] Rat oncogenic astroblastoma C6 cultural cell Mb Mb-bound 5'-nucleotidase Inhibition with ConA ConA+ [667]

3.1.3.(5). 5'-Nucleotidase-like Murine liver microsomes 5'-Nucleotidase specific for orotidine-5'-phosphate, 53kDa Biobeads-SM2—LCA-Sepharose4B** LCA- (227%, 9,7-f) [668]

3.1.3.(5). 5'-Nucleotidase-like Glycine max L.(soybean sedds) 5'-UMP-specific phosphatase 45kDa, forms I and II DEAE-Toyopearl650M(I+II)—DEAE- Toyopearl650M—ConA-Sepharose— ConA+ (desorption with Toyopearl-HW55 0,1M MM; 39%,123-f, for I; 72%, 9.4-f, for II) [669] 3.1.3.6. 3'-Nucleotidase

Protozoans Leishmania donovani strain S1 (leishmanial promastigote surface) 3'-Nucleotidase 43kDa (SDS-PAGE) solubilized with OG DEAE-Sephacel—ConA-Sepharose**— ConA+ (desorption with MonoS*—Superose12* 0,3M MM; 70%, 9,0-f) [670]

Asn-glycans: 3kDa in 43 kDa N-Glycanase+ (43—40kDa)

[671] 3'-Nucleotidase major form 42kDa, and minor form 38kDa, solubilized with OG DEAE-Sephacel—ConA-Sepharose— ConA+ (desorption with Q-Sepharose* 0,3M MM) [672]

Leishmania donovani strain 15 (destroyed cells) 3'-Nucleotidase 38kDa, pI 5,8, solubilized with TX100 ConA-Sepharose—DEAE-Sephacel**—IEF ConA+ (complete binding; desorption with 50mM MM; 67%, 2,1-f) [673] Higher plant Pisum sativum L.(pea germinating seeds) 3'-Nucleotidase with SU 30+24kDa ConA-Sepharose—DEAEC—SephadexG100 ConA+ (desorption with 1M Glc; separation from phosphatases) [674] Carbohydr.: 20% [675]

3.1.3.8. 3-Phytase Aspergillus ficuum NRRL 3135 (fungal CF) Enzyme 90kDa(Biogel-P200), or PAS+ (for native or EndoH- 85+100kDa (SDS-PAGE), pI 4,5 treated enzyme) SP-TrisacrylM—DEAE-TrisacrylM— Chromatofoc** Asn-glycans of oligoMan type: 11 or EndoH+ ([85+100]—76kDa) 24%, for 85 or 100kDa, respectively [676]

3.1.3.16. Phosphoprotein phosphatase Bovine spleen Enzyme Phosphocellulose**—Phosphocellulose— SephadexG75—ConA-Sepharose4B ConA+ (desorption with 0,5M MM; 67%, 7,8-f) [677] 3.1.3.31. Nucleotidase Leishmania tropica (leishmanial promastigote surface) Nucleotidase solubilized with TX100 CMC—SephacrylS300—ConA- ConA+ (desorption with Sepharose—PAGE sugar) Inhibition of surface enzyme with ConA ConA+ [678]

3.1.3.48. Protein-tyrosine-phosphatase

Insect Spodoptera frugiperda cells SF9 Human brain protein-tyrosine- phosphatase expressed in cells SF9, forms 98 and 114kDa Asn-glycans of complex type: 16kDa EndoF+ (114—98kDa), in 114kDa EndoH- [679]

54

Higher plant Triticum vulgare (wheat seedlings) Monomeric protein-tyrosine- phosphatase 35kDa Phosphocellulose—DEAEC—Phospho- cellulose—CM-Sephadex**—ConA- ConA+ (desorption with 30 Sepharose mM MM using gradient 0-0,15M sugar; 21%, 3,3-f) [680] Mammals Human cells D98/AH-2 (ATCC CCL 18.3) (mitotic cell extract) Nuclear protein-tyrosine-phosphatase 65kDa DEAE-Sepharose—Ab-sorbent, WGA-agarose WGA+(desorption with 0,1M GlcNAc) [681] Ab-precipitate —Blot —WGA WGA+, WGA- (for alkaline- treated blot) Carbohydr.: clusters of O-GlcNAc [682]

Human T-lymphocyte Mb Protein-tyrosine-phosphatase of CD45- antigen (GP 220+205+190+180kDa) associated with SU of PHA (33kDa) [683]

Mixture of glycans + + + + Lectin-agarose (sequential PHA-E4 , ConA , DSA , AAL , chromatography) SNA+ (separation of glycans)

Mainly bi-, tri-, and tetrantennary Asn-glycans of complex type [684]

Mammalian B-lymphocyte Mb Phosphotyrosine phosphatase isozymes of antigen CD45 Binding to CD22 (Sia-specific soluble CD22+ recombinant antigen from monkey COS cells) [685]

3.1.3.60. Phosphoenolpyruvate phosphatase Brassica nigra (black mustard, suspension cells from leaf petioles) Cell monomeric enzyme 55kDa(SDS- PAS+ PAGE),pI 4.5 S-Sepharose(*)**—Cu2+-chelating- Sepharose —Superose12*, ConA-agarose ConA+ (desorption with gradient 0-0,5M MM; 67%, 6,0-f) [549,686] 3.1.3.(64). Polyphosphoinositide phosphatase Rat brain Mb Monomeric enzyme 85-90kDa solubilized with TX100 DEAEC—Blue-agarose—Zn2+-chelating-

agarose—SephacrylS300—Heparin- agarose— (ConA-agarose—WGA- ConA-WGA+ (desorption with agarose as coupled column)** 50mM GlcNAc; 48%, 8,9-f) [687]

3.1.4.1. Phosphodiesterase I

Lower plants Aspergillus niger FS-44 (fungal micelium) 5'-Phosphodiesterase associated with cyclic-ribonucleotide phosphomutase, 62kDa (SDS-PAGE, SephacrylS200), pI 4.8 DEAE-Toyopearl**—SP-Toyopearl— 2',5'ADP-Sepharose Asn-glycans: 6kDa in 62kDa N-Glycanase+(62—56kDa) [688] Fusarium moniliforme (fungus) Monomeric phosphodiesterase-phospho- monoesterase 100kDa, 4 forms PAS+ ConA-Sepharose, IEF—ConA-Sepharose ConA+ (desorption with 50 mM MM using gradient 0- 0,5M sugar;similarity of all forms) Carbohydr.: 35.8% hexoses, 3.6% GlcNAc [689]

Saccharomyces cerevisiae IBPhM-366 (yeast Mb) Mb-bound pyrophosphatase 120kDa, SU 41 kDa,solubilized with TX100, and soluble enzyme form 82kDa, SU 28kDa SephacrylS300—MonoQ* Carbohydr. in both forms [690]

Higher plant Oryza sativa L. cv.Nipponkai (rice embryonal cells in culture) Golgi-Mb-bound nucleoside diphosphatase 200kDa, SU 55kDa, pI 7.5, solubilized with TX100 ConA-Sepharose4B**—DEAE-Toyopearl ConA-(309%, 4,7-f) —EAH-Sepharose4B—Cellulofine-GCL- 2000sf [691]

Mammals Bovine liver plasma Mb Alkaline nucleotide phosphodiesterase associated with threonine-specific protein kinase, 260kDa, SU 130kDa WGA-Sepharose WGA+ [692]

Human placenta Phosphodiesterase-I associated with nucleotide-pyrophosphatase, 520kDa, SU 130kDa

56

Affigel-Blue**—AMP-agarose—WGA- WGA+ (desorption with Sepharose6MB—ADP-agarose— 0,4M GlcNAc; 96%, 2-f) SepharoseCL6B—DEAEC [693]

Human plasma 5'-Nucleotide-phosphodiesterase as platelet aggregating factor 230kDa, SU 66+45+16kDa SephacrylS200—DEAEC**—Sephacryl- S300—SephacrylS300, ConA-Sepharose ConA+ (desorption with 25 mM MM; inactivation of 90% in eluate) SDS-PAGE—ConA-125I ConA+ (66+45kDa) [694]

Murine plasmacytoma MOPC 315 cell Mb Phosphodiesterase-I associated with murine plasma cell differentiation antigen PC1, 115kDa (SDS-PAGE) Asn-glycans: 12kDa in 115kDa N-Glycanase+ (115—103kDa) [695]

Rat liver Mb Phosphodiesterase-I 125kDa ConA-Sepharose—5'AMP-Sepharose— ConA+ (desorption with DEAE-SepharoseCL6B—SephacrylS400, 0,5M MM), LCA-Sepharose LCA+ [696]

(3.1.4.1.) Activating factor of phosphodiesterase-I Frog retinal rod outed segment Mb Detergent-solubilized opsin containing protein activator of phosphodiesterase-I ConA-Sepharose ConA+ (desorption with 0,2M MM) [697] 3.1.4.3. Higher plant Arachis hypogaea L. (peanut seeds) Inositol phospholipid-specific phospholipaseC DEAEC**—Octyl-Sepharose—ConA- ConA+ (desorption with Sepharose 0,5M MM; 49%, 5,9-f) [698] Mammals Mammalian tissue vesicular Mb Phospholipase-C forms 39 and 37kDa (SDS-PAGE) SDS-PAGE—Blot—Lectin ConA+, WGA- [699]

Rat brain Mb Thyrotropin-realising-hormone-degrading phospholipidase type-III 230kDa, SU 116kDa, solubilized with trypsin Q-Sepharose(*)**—Octyl-Sepharose— LCA-Sepharose—Blue-Sepharose—TSKgel- LCA+ (desorption with DEAE-5PW*—TSKgelG3000SW*, 0,7M MG; 89%, 26-f) Lectin-agarose ConA+ or WGA+ (strong binding)

SDS-PAGE—Blot—Lectin-biotin ConA+, PHA-E+, WGA+ Asn-glycans: 19kDa in 116kDa EndoF+ (116—97kDa) [700]

3.1.4.4. Higher plant Arachis hypogaea L. (peanut seeds) Phospholipase D DEAEC—Octyl-Sepharose—ConA- ConA+ (desorption with Sepharose** 0,5M MM; 42%, 14-f) [698] Mammals Bovine serum PhospholipaseD 200kDa, SU 100kDa, pI 5,6 Q-Agarose*—SephacrylS300—WGA- WGA+ (desorption with Sepharose**—HA—Zn2+-chelating- GlcNAc;71%, 6,2-f) sorbent—MonoQ* Ab-Sepharose**—WGA-Sepharose— WGA+ (62%,10-f) MonoQ* [701]

Human plasma Monomeric phospholipaseD 110kDa MonoQ*—Phenyl-Glass-Pac5PW(*)**— WGA+ (complete binding; WGA-Sepharose6MB—MonoQ* desorption with 0,45M GlcNAc; 27%, 2,7-f) [702] 3.1.4.12. Sphingomyelin phospho- diesterase

Bovine seminal plasma Acid sphingomyelinase 160kDa, SU 60kDa SephacrylS300—Q-Sepharose(*)**— S-Sepharose*—ConA-Sepharose— ConA+ (74%, 3,0-f) Superose-6* [703]

Human brain Sphingomyelinase forms: A (150kDa) and B (60kDa) ConA-Sepharose4B**—SephadexG200 ConA+(complete binding of A and B; desorption with 0,75M MM; 94%, 14-f [704] Sphingomyelinase 170-210kDa, SU 67kDa ConA-Sepharose**—CMC—HA— ConA+ (desorption with BiogelA1.5m MM; 63%, 54-f) [705]

Human brain and liver from patients with Nymak-Peak disease Sphingomyelinase forms I and II ConA-Sepharose—SephadexG200— ConA+ DEAEC—CMC [706]

Human liver Acid sphingomyelinase of extract Lectin-agarose ConA+ (binding of 82%; desorption of 69%), WGA+ (binding of 41%; desorption of 21%),

58

DBA+ (12%), RCA-I+(5%), UEA-I+(12%), SBA- [707] Neutral sphingomielinase Lectin-agarose ConA+ (63%), DBA+ (29%), WGA+ (32%), UEA-I+ (7%), RCA-I-, SBA- [707] Human placenta Sphingomyelinase forms pI 6,2-7,5, solubilized with TX100 ConA-Sepharose**—Sepharose6B—CM- ConA+ (desorption with Sepharose—IEF 0,5M MM; 42%, 68-f) [708]

Sphingomyelinase with SU 89,1 kDa ConA-Sepharose**—Sphingosylphospho- ConA+ (107%,108-f) choline-Sepharose; Octyl-, Hexyl-, and Blue-Sepharose [709]

Sphingomyelinase 123kDa, SU 62kDa, pI 7,4, solubilized with OG ConA-Sepharose**—Sphingosylphospho- ConA+ (desorption with choline-Sepharose—Hexyl-agarose— 0,5M MM; 100%, 21-f) MonoP* [710]

Sphingomyelinase with SU 70kDa solubilized with NP40 ConA-Sepharose—Butyl-agarose**— ConA+ (41%, 1,6-f) Octyl-agarose—Blue-Sepharose— Sphingosylphosphocholine-Sepharose

Asn-glycans: 10kDa in 70kDa EndoF+(70—60kDa) [711]

Human urine Sphingomyelinase 200kDa, SU 70kDa Octyl-Sepharose—ConA-Sepharose— ConA+(washing with 10mM Blue-Sepharose**—DEAEC MG followed by desorption with 0,75M MG; 57%, 5,9-f) [712,713] ConA-Sepharose—Octyl-Sepharose ConA+ (desorption with MM) [714] Rat liver lysosomes Heterotrimeric sphingomyelinase 220kDa DEAEC—Octyl-Sepharose—Sephacryl- S300—ConA-Sepharose—CMC** ConA+ (binding of 90%; desorption with 0,75M MM; 55%, 8,9-f [715] (3.1.4.12.) Protein activator of sphingomyelinase Human Gaucher spleen fibroblasts Sphingomyelinase activator forms 6 and 3,5 kDa (SDS-PAGE) ConA-Sepharose—HA—Octyl-Sepharose— ConA- (3,5 kDa),ConA+ DEAE-Sephacel (6kDa; desorption with 0,75M MM) [714] 3.1.4.16. 2',3'-Cyclic-nucleotide 2'-phosphodiesterase Solanum tuberosum (potato tubers)

2',3'-Cyclic-nucleotide 2'-phospho- diesterase associated with nucleotide pyrophosphatase PAS+ HA—HA—SephacrylS300**—Sephacryl- S300, ConA-Sepharose ConA+ [716]

3.1.4.17. 3',5'-Cyclic-nucleotide phosphodiesterase Dictyostelium discoideum (slime mold) Extracellular enzyme forms 150-200kDa, pI 5,0, and 55kDa, pI 7,5-9,0, SU 48+50kDa (SDS-PAGE) SephacrylS200—Affigel-Blue—ConA- ConA+ (desorption of 64% Sepharose4B—Affigel-Blue—Ultrogel- with 50mM MM, for 55kDa) AcA34 SephacrylS200—DEAEC—ConA- ConA+ (150-200kDa; Sepharose4B—IE—Sepharose6B 95%, 1,6-f) [717]

Cell cCMP-hydrolase ConA-Sepharose ConA- (separation from the bound cAMP-hydrolase) [718] (3.1.4.17.) Protein inhibitor of cyclic nucleotide phosphodiesterase Dictyostelium discoideum axenic strain AX3 (slime mold, CF of early stationary phase culture) cAMP-hydrolase inhibitor 47kDa (SDS-PAGE) SephacrylS200—ConA-Sepharose4B— ConA+(89%,19-f), DEAEC—BiogelA1.5m ConA- (perjodate-treated inhibitor) [719] 3.1.4.35. 3',5'-Cyclic-GMP phospho- diesterase

Lower plant Dictyostelium discoideum (slime mold) cGMP-dependent cyclic-GMP-specific phosphodiesterase ConA-Sepharose ConA- (separation from the bound cAMP-hydrolase) [718] Mammals Bovine and monkey retina Extracellular cGMP-phosphodiesterase (interphotoreceptor matrix enzyme) 350kDa, SU 47+45kDa BiogelA1.5m—ConA-Sepharose4B— ConA-(separation from Protamine-agarose—TSKgelG3000SW* alpha2-macroglobulin SU 114kDa and inter-photoreceptor retinol-binding protein) [720] 3.1.4.37. 2',3'-Cyclic-nucleotide 3'-phosphodiesterase Bovine brain white substance Enzyme with SU 56+53kDa Lectin-agarose WGA+ (desorption of 50% with 0,1M GlcNAc), RCA-I+, ConA+ (desorption with 1M

60

KCl), UEA-I- [721] 3.1.4.40. CMP-N-acetylneuraminate phosphodiesterase Rat liver Mb CMP-sialate hydrolase 500kDa, SU 50-55kDa ConA-Sepharose — 5'-AMP-Sepharose — ConA+ (desorption with 0,4 DEAE-SepharoseCL6B— SephacrylS400, M MM), LCA+ (less effective LCA-Sepharose than ConA) [696]

3.1.4.45. Acetylglucosamine-1-phospho- diester N-acetylglucosaminidase Bovine liver Mb N-Acetylglucosamine-1-phosphodiester beta-N-acetylglucosaminidase with SU 129+121kDa(SDS-PAGE) solubilized with TX100 DEAEC**—ConA-Sepharose—WGA- ConA+ (desorption with 0,1 Sepharose—Superose6*—SDS-PAGE M MM; 50%, 2,1-f), WGA+ (desorption with 0,1M GlcNAc; 21%, 2,7-f), ConA+WGA+ (10%, 6,0-f) [722] Rat liver Mb beta-N-Acetylglucosaminyl phospho- diesterase solubilized with TX100 DEAEC—Heparin-Sepharose**—ConA- ConA+ (desorption with Sepharose 0,5M MM; 61%, 4,4-f) [723] 3.1.6.1.

Invertebrate Eimeria tenella (hen parasitic helminth, oocytes and sporozoits) Arylsulfatase 49kDa (SephacrylS200) Lectin-Sepharose4B ConA-, RCA-I- [724]

Birds Hen brain Arylsulfatase of heat-treated extract ConA-precipitation ConA+ [725]

Hen caecum Arylsulfatase 97kDa (SephacrylS200) ConA-Sepharose**—cAMP-Sepharose ConA+(desorption with 0.5M MM;87%,12-f) Carbohydr. PAS+ [724]

3.1.6.2. Steryl-

Bovine liver ArylsulfataseC forms pI 4,5 and 7,0 Phenyl-Sepharose—DEAE-Sephacel— ConA-Sepharose**—SephacrylS500 ConA+ (desorption with 0,5 M MG; 29%, 4,7-f) [726] Dog liver Homogenic arylsulfataseC (PAGE)

Phenyl-Sepharose—DEAE-Sephacel— ConA-Sepharose**—SephacrylS500 ConA+ (desorption with 0,5 M MG; 94%, 5,6-f) [726]

Human liver ArylsulfataseC 200kDa, pI 6,5 DEAE-Sephacel—ConA-Sepharose**— ConA+ (desorption with 0,5 DEAE-Sephacel—DEAE-Sephacel— M MM; 70%, 15-f) Cellogel [727]

Human placenta ArylsulfataseC 183kDa, pI 6,5 DEAE-Sephacel**—ConA-Sepharose— ConA+ (desorption with 0,5 DEAE-Sephacel—DEAE-Sephacel— M MM; 95%, 6,6-f) Cellogel [727]

Arylsulfatase 238kDa, SU 78kDa DEAEC—ConA-Sepharose**—Biogel- ConA+(desorption with 0.5 A1.5m M MM; 62%, 10-f) [728]

Human placental microsomal Mb 390kDa solubilized with cholate derivative SepharoseCL6B—ConA-Sepharose4B ConA+ (complete binding; desorption with 0,4M MM) Carbohydr.moiety important for beta-N-Acetylglucosaminidase+, activity alpha-Mannosidase+, EndoH-, Lysozyme+, Sialidase- [729] ArylsulfataseC 130kDa BiogelA1.5m—DEAEC—PAGE—ConA- ConA+(separation from from Sepharose unbound sulfatase) [730]

Rat liver Homogenic arylsulfatase (PAGE) Phenyl-Sepharose—DEAE-Sephacel— ConA-Sepharose**—SephacrylS500 ConA+ (desorption with 0,5 M MG; 84%, 8,9-f) [726] Microsomal arylsulfataseC 280kDa, SU 72kDa, pI 8,1, solubilized with TX100 DEAEC**—DOC-Sepharose—Hexyl- Sepharose—SephadexG200—Hexyl- Sepharose, ConA-Sepharose ConA+ (strong binding;low desorption with MM) Carbohydr. (M/M): 20 Man, 2,6 Glc, 1,1 Gal, 4,9 GlcNAc; Asn-glycans of oligoMan type EndoH+ [731]

3.1.6.4. N-Acetylgalactosamine-6- sulfatase Human placenta Galactose-6-sulfate sulfatase 100kDa ConA-Sepharose—DEAEC—Sephacryl- ConA+ S200—Prepar.PAGE [732]

N-Acetylgalactosamine-6-sulfate

62 sulfatase 120kDa, SU 40+15kDa PAS+ (for 15kDa) ConA-Sepharose**—DEAEC—Ultrogel- ConA+ (desorption with 0,5 AcA34—MonoP*—MonoQ* M MM; 90%, 64-f) [733]

3.1.6.8. Cerebroside-sulfatase

Invertebrates Helix pomatia (garden snail) ArylsulfataseA 90kDa (PAGE, pH 7,5), or 100kDa (PAGE, pH 4,5) ConA-Sepharose—DEAE-Sepharose— ConA+ SephacrylS200 [734]

Patella sp.(limpet) ArylsulfataseA 125kDa (PAGE, pH 7,5), or 145kDa (PAGE, pH 4,5) ConA-Sepharose—DEAE-Sepharose— ConA+ Sephacryl-S200 [734]

Fishes Salmon liver ArylsulfataseA 180kDa (PAGE, pH 7,5), or 189kDa (PAGE,pH 4,5) ConA-Sepharose—DEAE-Sepharose ConA+ SephacrylS200 [734]

Shark liver ArylsulfataseA 180kDa (PAGE, pH 7,5), or 184kDa (PAGE, pH 4,5) ConA-Sepharose—DEAE-Sepharose ConA+ SephacrylS200 [734]

Trout liver (Salmo sp.) ArylsulfataseA 180kDa (PAGE, pH 7,5), or 189kDa (PAGE, pH 4,5) ConA-Sepharose —DEAE-Sepharose ConA+ SephacrylS200 [734]

Birds Hen liver ArylsulfataseA 166kDa (PAGE, pH 7,5 or 4,5) ConA-Sepharose—DEAE-Sepharose ConA+ SephacrylS200 [734]

Pigeon liver ArylsulfataseA 180kDa (PAGE, pH 7,5) or 182kDa (PAGE, pH 4,5) ConA-Sepharose—DEAE-Sepharose ConA+ SephacrylS200 [734]

Reptiles Turtle liver ArylsulfataseA 166kDa (PAGE, pH 7,5), or 197kDa (PAGE, pH 4,5) ConA-Sepharose—DEAE-Sepharose— ConA+

SephacrylS200 [734]

Mammals Bovine liver Ascorbate-2-sulfate sulfohydrolase forms 59 and 52kDa (PAGE with urea) HA—SephadexA50—HA—Prepar.ultra- centrifugion, ConA-Sepharose ConA+ (irreversible binding: immobilization) [735] ArylsulfataseA 182kDa (PAGE, pH 7,5), or 324kDa (PAGE, pH 4,5) ConA-Sepharose—DEAE-Sepharose— ConA+ SephacrylS200(or Monomeric-enzyme- Affigel10) [734]

Purified sulfataseA modified with nitrocathechol sulfate ConA-Sepharose ConA+ (desorption with 0,1 M MG or MM;separation from unbound enzyme reaction products) [736] Feline liver ArylsulfataseA 180kDa (PAGE, pH 7,5), or 300kDa(PAGE,pH 4.5) ConA-Sepharose—DEAE-Sepharose— ConA+ SephacrylS200(or Monomeric-enzyme- Affigel10) [734]

Dog liver ArylsulfataseA 182kDa (PAGE, pH 7,5), or 297kDa (PAGE, pH 4,5) ConA-Sepharose—DEAE-Sepharose— ConA+ SephacrylS200(or Monomeric-enzyme- Affigel10) [734]

Horse blood leukocytes ArylsulfataseA with pI 4,3 ConA-Sepharose—Blue-Sepharose— ConA+ (desorption with 0,5 DEAEC—Phenyl-Sepharose* M MG; 60%, 6,0-f) [737]

Horse liver ArylsulfataseA 180kDa (PAGE, pH 7,5), or 348kDa (PAGE, pH 4,5) ConA-Sepharose—DEAE-Sepharose— ConA+ SephacrylS200(or Monomeric-enzyme- Affigel10) [734]

Human cell lines HeLa, PC-3SF12, WI-38 ArylsulfataseA 64kDa (WI-38), 67kDa (HeLa), or 69kDa (PC-3SF12), SDS-PAGE ConA-Sepharose—Ab-precipitation ConA+ Carbohydr.: 7, 8, 12, or 10%, for 64, 67, or 69kDa, respectively; Asn-glycans of oligoMan type: 8 kDa in EndoH+ ([64+67]—59kDa; 67 kDa, 7 kDa in 69 kDa 69—62kDa) [738]

64

Human liver ArylsulfataseA 150kDa (PAGE, pH 7,5), or 330kDa (PAGE, pH 4,5) ConA-Sepharose—DEAE-Sepharose— ConA+ SephacrylS200(or Monomeric-enzyme- Affigel10) [734]

Human lung ArylsulfataseA 100kDa, SU 58kDa, 6 forms pI 4,6-5,1 DEAE-Sephacel**—SephadexG200— ConA-Sepharose—SephadexG200 ConA+(78%, 2.9-f) Carbohydr.: Sia; Asn-glycans of Arthrobacter sialidase+— oligoMan type EndoH+ (sequential treatment) [739] Human placental microsomes Mb-bound estrogen-sulfatase form 390kDa or solubilized with TX100 form 270kDa SepharoseCL6B—ConA-Sepharose ConA+ (complete binding; desorption with 0,4M MM) Carbohydr. moiety important for beta-N-Acetylglucosaminidase+, activity alpha-Mannosidase+ [729]

Human platelets of normal and alcoholic patients ArylsulfataseA with SU 62+57kDa DEAE-Sepharose—SephacrylS300, Lectin-Sepharose4B (LCA, PHA-E, Lectin+ (separation of PHA-L, RCA-I, RCA-II, SBA, WGA) isozymes IIIa, IIIb, and IVb, characteristical of alcoholic patients) Asn-glycans PNGaseF+ [740]

Human urine ArylsulfataseA with SU 69, 63, and 54,3kDa ConA-Sepharose**—DEAE-Sepharose— ConA+ (desorption with 0,3 Prepar.PAGE—SephacrylS300, M Man; 69%, 119-f) (Monomeric-rabbit-liver-arylsulfataseA)- Sepharose**—SephacrylS200 Carbohydr. EndoH-, PNGase- [741]

Monkey brain ArylsulfataseA of extract Lectin-Sepharose ConA+ (desorption with MM or MG), RCA-I- [742] Opossum liver ArylsulfataseA 132kDa (PAGE, pH 7,5), or 175kDa (PAGE, pH 4,5) ConA-Sepharose—DEAE-Sepharose— ConA+ SephacrylS200 [734]

Porcine liver ArylsulfataseA 160kDa (PAGE, pH 7,5), or 330kDa (PAGE, pH 4,5) ConA-Sepharose—DEAE-Sepharose— ConA+

SephacrylS200(or Monomeric-rabbit- liver-enzyme-Affigel10 [734]

Rabbit kidney cortex ArylsulfataseA SephadexG150—DEAEC**—Sephadex— ConA-Sepharose—SephadexG200 ConA+ (desorption with 0,6 M MM; 59%, 9,2-f) [743] Rabbit liver ArylsulfataseA 130kDa (PAGE, pH 7,5), or 330kDa (PAGE, pH 4,5) ConA-Sepharose—DEAE-Sepharose— ConA+ SephacrylS200(or Monomeric-enzyme- Affigel10) [734] ConA-Sepharose—Monomeric-enzyme- ConA+(desorption with Affigel10 0,5M Man) [744]

Rabbit seminal plasma ArylsulfataseA Precipitation with ConA ConA+ (complete; retention of 20% in precipitate) [745] Rat liver Ascorbate-2-sulfatase forms 65 and 104 kDa (SDS-PAGE) DEAE-Sephacell—ConA-Sepharose**— ConA+ (desorption with 1M SephacrylS300 MM; 50%, 7,3-f, for 65 kDa; 96%, 17-f, for 104kDa) [746] Carbohydr.: 10-22% hexoses [747]

Seal liver ArylsulfataseA 180kDa (PAGE, pH 7,5), or 330kDa (PAGE, pH 4,5) ConA-Sepharose—DEAE-Sepharose— ConA+ SephacrylS200(or Monomeric-enzyme- Affigel10) [734]

Sheep brain ArylsulfataseA 122kDa, SU 63kDa ConA-Sepharose**—DEAE-Sepharose— ConA+ (desorption with 0,5 SephadexG200(pH 5)—SephadexG200 M MG; 59%, 100-f), (pH 7.5), RCA-I-Sepharose4B RCA-I- Carbohydr.: 25% hexoses, 0,4% Sia [748]

Sheep liver ArylsulfataseA 180kDa (PAGE, pH 7,5), or 348kDa (PAGE, pH 4,5) ConA-Sepharose—DEAE-Sepharose— ConA+ SephacrylS200(or Monomeric-enzyme- Affigel10) [734]

Whale liver ArylsulfataseA 180kDa (PAGE, pH 7,5), or 340kDa (PAGE, pH 4,5) ConA-Sepharose—DEAE-Sepharose— ConA+ SephacrylS200(or Monomeric-enzyme-

66

Affigel10) [734]

(3.1.6.8.) Protein activator of galactosylceramide sulfatase Human liver 22kDa, SU 8kDa DEAEC—SephadexG100—SephadexG50 —Prepar.PAGE Carbohydr.: Man, Gal, GlcN [749]

3.1.6.12. N-Acetylgalactosamine 4-sulfatase

Feline liver Enzyme 166kDa, SU 41kDa, pI 7,8 DEAEC—ConA-Sepharose**—Blue- ConA+ (desorption with 1M Sepharose—CMC—SephadexG200— MG; 76%, 465-f) DEAEC—SephadexG200 [750]

Horse blood leukocyte lysosomes ArylsulfataseB forms pI 8,6 and 8,9 ConA-Sepharose**—Blue-Sepharose— ConA+ (desorption with CMC—SephadexG200 0,5M MG; 60%, 6,0-f) [737] Human liver Lysosomal N-Acetylgalactosamine 4-sulfate sulfatase 62kDa, pI 5,7 (ConA-Sepharose—Blue-agarose)**— ConA+ (complete binding; Chromatofoc—TSK-HW50S- desorption with MM), Fractogel—Cu2+-chelating-Sepharose — ConA—Blue (86%,104-f) Phenyl-Sepharose—TSKgelG3000SW*— Cu2+-chelating-Sepharose [751]

Enzyme 80kDa, SU 38kDa, pI 8,0 ConA-Sepharose**—Blue-Sepharose— ConA+ (desorption with CMC—SephadexG200—DEAEC— 1M MG; 70%, 263-f) SephadexG200 [750]

Enzyme 43kDa (UltrogelAcA34), or 57kDa (SDS-PAGE) ConA-Sepharose**—Blue-agarose— ConA+ (desorption with Blue-agarose—HA—Blue-agarose— 0,75M MM; 53%, 90-f) Cu2+-chelating-Sepharose, ConA-Sepharose—mAb(ASB-4.1)- Affigel10** [752]

Human placenta Intracellular N-acetylgalactosamine 4-sulfatase with SU 47kDa DEAEC—CMC—CMC—ConA-Sepharose ConA+ (desorption with —SephadexG100—SephadexG75—DEAEC 0,5M MM; 59-f) [753]

Ox liver N-Acetylgalactosamine 4-sulfate sulfataseB1 59kDa ConA-Sepharose—ConA-Sepharose ConA+ (desorption with

0,1-0,3M MM using gradient 0-0,4M MM; >80%, 10-f; separation of forms) Carbohydr.: GlcN [754]

Rabbit kidney cortex ArylsulfataseB SephadexG150—DEAEC—CMC—ConA- ConA+ (Desorption with Sepharose 0,6M MM; 47%, 37-f) [743]

Sheep brain N-Acetylgalactosamine 4-sulfatase 45kDa (SephadexG200) ConA-Sepharose**—Blue-dextran- ConA+ (desorption with precipitation—DEAEC 0,5M MG; 66%, 69-f) Carbohydr.: 11% hexoses, 0.4% Sia [755]

3.1.6.13. Iduronate-2-sulfatase

Human liver Enzyme forms A (42-65kDa, SU 42+14kDa), and B (SU 42+14kDa) (ConA-Sepharose—Blue-agarose)**— ConA+ (recyclic desorption Chromatofoc—TSK-HW50S-Fractogel— with 0,5M MM), Phenyl-Sepharose—TSKgelG3000SW* ConA—Blue (76%, 119-f) [756]

(ConA-Sepharose—Blue-agarose— ConA+ (desorption with Green-Sepharose) 0,5M MM; 92%, 39-f) [757]

Human placenta Monomeric iduronate sulfatase 80kDa ConA-Sepharose**—DEAE-BiogelA— ConA+ (93%, 53-f) PreparPAGE—Phenyl-Sepharose [758] Enzyme 80-100kDa, pI 4,5-4,8 DEAE-Sepharose—ConA-Sepharose**— ConA+ (desorption with DEAE-Sepharose—MonoQ*—MonoP* sugar; 73%, 87-f) [759]

3.1.6.14. N-Acetylglucosamine- 6-sulfatase

Bovine kidney Enzyme 58-60kDa (Superose-6*, SDS-PAGE) associated with novel endosulfatase activity SephacrylS200—DEAEC—CMC**—CMC —Lectin-Sepharose ConA+,WGA+,RCA-I- Asn-glycans [760]

Human liver N-Acetylglucosamine-6-sulfate sulfatase forms A (monomeric 75-78kDa, pI>9,5), B (75kDa, SU 48+32kDa, pI>9,5), C (94kDa, SU 48+45+32kDa, pI 5,8), and D (75kDa, pI 5,4) (ConA-Sepharose—Blue-agarose)**— ConA+ (complete binding Chromatofoc—Cu2+-chelating- of A and B; recyclic

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Sepharose—HA—Gel-fitration— desorption with MM), Cu2+-chelating-Sepharose ConA—Blue (210%,163-f) [761] 3.1.6.18. Glucuronate-2-sulfatase Human liver Enzyme 63kDa, SU 43kDa, pI>7,5 (ConA-Sepharose—Blue-agarose)**— ConA+ (desorption with (DEAE-Sephacel—Octyl-Sepharose)— 0,75M MM), CM-Sepharose—TSKgelG3000SW* ConA—Blue (64%, 853-f) [762] 3.1.15.1. Venom I Crotalus adamanteus (snake venom) Enzyme ConA-Sepharose (Calbiochem, USA) ConA+ (binding of 80% without desorption by 0,1M MM;separation from nonspecific phosphatases and endonuclease) [763] Enzyme 120kDa ConA-Sepharose**—BiogelP150— ConA+ (washing with 50mM NADP-agarose MM; desorption with 0,3M MM; 47%, 10-f; separation from nonspecific phosphatases,endonuclease and 10-12% 5'-exonuclease eluted by 50mM MM) [764] 3.1.16.1. Spleen exonuclease Bovine spleen Phosphodiesterase (5'-nuclease) 98kDa Ecteola-cellulose—ConA- ConA+ (desorption with Sepharose**—BiogelP150 0,1M MM; 78%, 6,0-f; separation from acid phosphatase and DNAse) [765] 3.1.21.1. (DNAse I)

Insect Drosophila melanogaster (fruit fly embrios) Lysosomal DNAse-I 32kDa (SDS-PAGE) HA—S-Sepharose*—Q-Sepharose*— Aminopentyl-agarose—Superose12*— ConA-Sepharose ConA+ (desorption with 0,6M MM; 43%, 2.8-f) [766] Mammals Bovine pancreas DNAses A, B, C, and D ConA-Sepharose ConA- (A and B), ConA+ (complete binding of C and D; desorption with 0,5M Glc) [767,768] Carbohydr.(M/M): 5-6 Man : 2 GlcN; Sia in A and B [768-770]

DNAse Type III (Sigma, USA) including 4 major forms and few minor forms (2-dimential PAGE)

PAGE—Blot—Lectin-peroxidase ConA+, WGA+, RCA-I+ (for Sialidase-trerated enzyme) Asn-glycans mainly of oligoMan type and/or hybrid type [771]

Bovine spleen Acid DNAse Ecteola-cellulose—ConA-Sepharose ConA-(separation from the bound 3'-nuclease) [765] Guinea pig epidermis Neutral 33kDa, pI 5,2 DEAEC**—SephadexG100—ConA- ConA+ (desorption with Sepharose 1M MG; 29%, 2,4-f) [772]

Human urine DNAse from the man of 46 years DEAE-Sepharose**—Phenyl-Sepharose— HA—Elastin-Celite—ConA-agarose— ConA+(desorption with SephadexG75 1,5M Glc; 70%, 3-f; separation) from unbound proteases) Carbohydr.(M/M): 10 Man, 7 Gal, 1 Fuc, 6 GlcN, 2 Sia [770]

Ovine pancreas DNAse forms IA, IB, IIA, and IIB (monomeric II 31kDa, SDS-PAGE) CMC—ConA-Sepharose—CMC—Sephadex- ConA- (IA, IB), ConA+ G100 (complete binding of IIA, IIB) Carbohydr.: absence of Sia in all forms [768]

Rat small intestinal mucosa Neutral endoDNAse 32kDa, pI 4,7 DEAEC—Phosphocellulose**—Sephadex- G100—ConA-Sepharose ConA+ (desorption with 1M MG; 14%, 1,6-f) [773] (3.1.21.1.) Factor increasing DNAse-I inhibition with G- or F-actin Human plasma Actin-depolymerizing factor (brevin or gelsolin) 90kDa, pI 6,0-6,5 DEAEC—Blue-Sepharose—ConA- ConA- (separation from Sepharose contaminating GP) [774]

F-actin-depolymerizing factor 93kDa (SDS-PAGE), pI 5,84 DEAE-Sepharose—Chromatofoc**— ConA-Sepharose ConA- (separation from contaminating Ig; 113%, 1,3-f) [775] 3.1.27.1. Ribonuclease T2 (RNAse T2) Aspergillus niger (fungus) RNAse T2 36 or 29kDa 5'-Adenylate-Sepharose, ConA-Sepharose (for purification) ConA+ [776,777]

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ConA-Sepharose (for prepering of ConA+ (retention of 70% biocatalyst) in immobilized form) [778] Carbohydr.(for 29kDa): 7.9% hexoses [777] For 36kDa (mkg/mg protein): 84 Man, 14 Gal, 12 Glc, 2.9 Xyl [776] Glycosylation sites: Asn15,76,239 [777]

RNAse T2L 80kDa including 5 isoforms Carbohydr.: Xyl, Man, Gal, Glc (relative contents depending on type of isoform) [776]

3.1.27.(1). RNAse T2-like Insect cell culture GP-E2 RNAse of classical swine fever virus (CSFV) expressed in insect cells [776a]

3.1.27.5.

Bovine pancreas RNAses A and B 2'(3')-CMP-Sepharose4B—ConA- ConA- (RNAseA), ConA+ Sepharose—CMP-Sepharose (RNAseB; desorption with 0,5M MG) [779] RNAse B (Types XII, XIII, Sigma, USA) ConA-Sepharose ConA+ (desorption with 0,5 M MG or 0,2M MM; separation from unbound RNAseA as impurity) [779-784] ConA-Sepharose—WGA-Sepharose ConA+WGA+ (binding of 35%) [785] RNAse (Reanal, Hungary) ConA-Glc-Spheron-H1000 (Chemapol, ConA+(separation of Prague) forms A and B) [786]

Purified RNAseB SDS-PAGE—Blot—Lectin-biotin (or ConA+, sWGA+, lectin-peroxidase) ConA- or WGA- (N-glycanase- treated enzyme) [787,788] Native RNAseB Affinity to lectins ConA+, hMBP- [789] RNAseB-Sepharose Affinity to lectins ConA+, hlMBP+, rlMBP+ [783,790] + Asn-glycans (RNAse B): ManGlcNAc2 alpha-Mannosidase (form1), Man5-8GlcNAc2 (form2), (form2) Man4GlcNAc2 (form3) [780] Fine structure of Asn-glycans of oligoMan type, for 5 enzyme glycoforms [791-793]

Nonenzymatic glycosylation of RNAseA [794,795]

Porcine pancreas RNAses A and B ConA-Sepharose4B ConA+ (RNAseB, separation from RNAseA) [782]

3.1.27.-. Ribonucleases

Lower plant Trichoderma viride (fungus) RNAse Trv with SU 34,5+30+27,5, pI 3,9, isolated from preparation "Cellulase T-AP"(Amano Pharm.Co., Japan) DEAE-Sephadex—SephadexG50—DEAE- Sephadex—DEAE—Sephadex—DEAEC— DEAE-Toyopearl650M—2'5'ADP-Sepharose Carbohydr.(M/M): 2.1% hexoses, 2.1 GlcNAc; Asn-glycans: 7kD in 34 kDa, 3kDa in 30 kDa EndoF+ ([34+30+27]—27kDa) [796] Higher plants Nicotiana alata (flower style extract) RNAses (GP "S") as style self- incompatibility gene products [797]

Nicotiana tabacum (cell suspension culture) RNAse NADP-Sepharose—ConA-Sepharose ConA-(separation from nuclease-I) [798] Vicia faba L. ssp. minor var. "Nadwislanski"(broad bean seedlings) spliting yeast rRNA and polyU-specific, 35kDa (Sephadex- G100), pI 7,7+6,7+5,3+4,4+3,8 SephadexG25—SephadexG100—DEAE- Sephadex**—ConA-Sepharose ConA+ (desorption of 92% with 50mM MM) [799] Mammals Human HeLa cell lysosomes Acid RNAse SephadexG100**—DEAEC—ConA-agarose ConA+ (strong binding; desorption of 20% with 0,5M MM; additional desorption with 0,75M MM next day; 37%, 1,7-f) [800] Human seminal plasma RNAse forms I (78kDa), II (16kDa, pI 9,9), III (13,3kDa, pI 11,0), and IV (5kDa, pI 4,8) ConA-Sepharose4B—DEAEC(or phospho- ConA+(all forms; desorption cellulose)— Agarose-5'-(4-aminophenyl- with gradient 0,1-0,5M MM; phospho)uridyl-2'(3')-phosphate— 24%, 9,6-f, for I and II; SephadexG75(or G100) 34%, 17-f, for III and IV) [801] Human urine RNAse forms Ul (large) and Us (short) Lectin-Sepharose ConA+, LCA+, AAL+, RCA-I+, WGA+ (separation of forms) [802] Carbohydr.: 28,5% (Ul), 5% (Us) Mixture of glycans

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Lectin-Sepharose (in combination with AAL+ and ConA+ (separation exoglycosidase- and endo-beta- of glycans) galactosidase-treatment) [803]

RNAse-1 16kDa (gel-filtration), or 17.5 kDa (SDS-PAGE) specific to pyrimidin Carbohydr.: 4,6% hexoses (Man, Fuc), 4,7% hexosamine (GlcN) [804]

Purified RNAse (128aar) similar to enzyme of seminal plasma Carbohydr.(M/M): 45 (Fuc, Man, Gal, GlcN, Sia, Xyl, Glc); Glycans of complex type at Asn34,76,88 [805]

3.1.30.1. Aspergillus Aspergillus oryzae (fungus) Nuclease-S1 32kDa (SDS-PAGE), pI 4,2, of commercial preparation (Sankyo Co. Ltd., Tokyo) SP-Sephadex—ConA-Sepharose— ConA+ (desorption with SephadexG100 0,4-0,8M MM using gradient 0-1,0 M MM), ConA- (EndoH-treated enzyme) [806]

DEAEC—CMC—ConA-Sepharose ConA+ (immobilization with retaining of 75%) [807] Asn-glycans of oligoMan-type [806]

3.1.30.(1). Single-stranded-RNA/DNA- specific nuclease Zinnia elegans L.(leaf mesophyll cells) Differentiation-specific nuclease 43kDa (Superose12*, SDS-PAGE) ConA-Sepharose ConA+ (desorption with 1M MM) LCA-Sepharose4B**—Phosphocellulose LCA+ (desorption with 0,1M MM; 70%, 72-f; separation from unbound RNAses) SDS-PAGE—Blot—ConA-biotin ConA+ (43kDa) [808]

3.1.30.(2). Serratia marcescens nuclease-like

Hordeum vulgare L. cv.Himalaya (barley grain aleurone granules) Purified enzyme 35kDa (SDS-PAGE) ConA-Sepharose ConA+ (desorption with 0,5M MM) Carbohydr.: 5,7%; Asn-glycans (2kDa in 33kDa) of oligoMan EndoH+ (35—33kDa) type [809]

Spinacia oleracea (spinach leaves)

Nuclease SP spliting single-strand/ duplex-DNA/RNA, 30,8 kDa, pI 7,7 DEAEC—Blue-Affigel—Poly(U)- Sepharose—SephadexG75—Poly(U)- Sepharose—ConA-Sepharose**— ConA+ (desorption with Superose12* 0,1M MM; 37%, 37-f) [810]

3.1.30.(2). Army worm nuclease Spodoptera litura (army worm larvae) Alkaline nuclease (nucleinate-nuclease) 18kDa(PAGE) Heparin-agarose, ConA-Sepharose ConA+(nearly complete binding;desorption with 1M Glc) Carbohydr.: 38% hexoses [811]

3.1.(30). Crotalus endonuclease Crotalus adamanteus (snake venom) Alkaline endonuclease ConA-Sepharose4B ConA- (separation from the bound phosphodiesterase) [763,764] 3.1.31.-. Nuclease-I hydrolizing single- strand DNA into 3'-mononucleotides Nicotiana tabacum (cell suspension culture, CF) Nuclease-I 35kDa, pI 5,2 and 5,6 NADP-Sepharose**—ConA-Sepharose4B— ConA+ (desorption with SephadexG75 0,3M MM; 80%, 1,1-f; separation from unbound EDTA-resistant RNAse) Carbohydr.: 9% hexoses [798]

Abbreviations ac acetyl(ated) aar amino acid residues AH aminohexyl approx. approximately Ara arabinose (m)Ab antibodies(monoclonal) blot blotting carbohydr. carbohydrates CF cultural fluid CHAPS 3-[3-(cholamidopropyl)dimethylammonoium]-1-propanesulfonate Chromatofoc chromatophocusing CMC carboxymethylcellulose DEAEC diethylaminoethylcellulose DOC deoxycholate DTAB dodecylmethylammonium bromide DTT dithiotreitol EDTA-Na ethylenediaminetetracetate-Na ELBA enzyme-lectin binding assay ELISA enzyme-linked immunosorbent assay

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EndoD,F,H,M endoglycosidases D,F,H,M f fold FF Fast Flow FITC fluoresceine isothiocyanate FPLC fast protein liquid chromatography Fuc fucose FucN fucosamine Gal galactose GalN galactosamine GalNAc N-acetylgalactosamine Galp galactopyranoside Glc glucose Glc-1-P glucose-1-phosphate GlcN glucosamine GlcNAc N-acetylglucosamine GP glycoprotein(s) Gp glycopeptide(s) hsc high speed centrifugion HA hydroxyapatite HPLC high pressure liquid chromatography HPLAC high pressure liquid affinity chromatography IEF isoelectric focusing Ig(A,E,G,M) immunoglobulins A,E,G,M IGF insulin growth factor IL interleukin(s) Lac lactose LAE lectin-affinity electrophoresis LIE lectin-immunoelectrophoresis LPS lipopolysaccharide(s) Mal maltose Man mannose ManN mannosaminyl Man-6-P mannose-6-phosphate Mb membrane(s) ME 2-mercaptoethanol (alpha-ME) Mel melibiose MG methyl-alpha-D-glucopyranoside MM methyl-alpha-D-mannopyranoside mol molecular MUF methyl-umbelliferyl NeuAc N-acetylneuraminic acid Neu5Ac N-acetyl-5-O-acetyl-neuraminic acid NP nitrophenyl pNP para-NP NP40 nonidet P-40 OG n-octylglucoside PAGE polyacrylamide gel electrophoresis PMSF p-phenylmethylsulfonyl ftoride

PNGase protein N-glycanase prepar. preparative PS polysaccharide(s) Rha rhamnose SDS sodium dodecyl sulfate Sia sialic acid STI soybean trypsin inhibitor SU subunit(s) TEAEC tetraethylaminoethylcellulose TFMSA trifluoromethane-sulfonic acid TX100 Triton X-100 Xyl xylose

Abbreviations and specificities of lectins Abbreviations, Full name and sources Specificitiesa

AAL Aleuria aurantia lectin(le.) Fuc-alpha-2(3/6) CEA Cuscuta europea ag. complex CFL Cucurbita ficifolia le. L(D)-Ara ConA ConcanavalinA (Canavalia ensiformis) Man-alpha-; Glc-alpha- sConA succinilated ConA(dimeric) Man-alpha-; Glc-alpha- ConG ConcanavalinG (Canavalia gladiata) Man-alpha-; Glc-alpha- HPA Helix pomatia ag. GalNAc-alpha-3GalNAc LCA Lens culinaris ag. Man-alpha-; Fuc-alpha-6 LCA-B isolectin B of LCA Man-alpha-; Fuc-alpha-6 LPA Limulus polyphemus ag. Sia LTA Lotus tetragonolobus ag. Fuc-alpha-2Gal PHA phytohemag.(Phaseolus vulgaris) complex PHA-E, E-PHA erythroag.of PHA complex PHA-E4 homotetrameric PHA-E complex PHA-L, L-PHA leukoag.of PHA complex PHA-M mucopolysaccharide form of PHA complex PHA-P protein form of PHA(PHA-E+PHA-L) complex PNA peanut ag.(Arachis hypogaea) Gal-1,3-GalNAc POA Pleurotus ostreatus ag. Fuc-alpha- ; Lac PSA Pisum sativum ag. Man-alpha-; Fuc-alpha-6 PVA Psathyrella velutina ag. GlcNAc PWM pokeweed mitogen (Phytolacca americana) RCA Ricinus communis ag. Gal-beta-3(4)GlcNAc RCA-I tetrameric RCA Gal-beta-3(4)GlcNAc SBA soybean ag. (Glycine max) GalNAc-1,3-Gal; Gal SEL Separatur le. complex SNA Sambucus nigra ag. Neu5Ac2-alpha-6Gal STA Solanum tuberosum ag. (GlcNAc-beta-4)2-3 TPA Tetragonolobus purpuratus ag. L-Fuc-alpha- UEA Ulex europaeus ag. complex UEA-I le.-I of UEA Fuc-alpha-2Gal VVA Vicia villosa ag. complex

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VVA-B4 isolectin B4 of VVA GalNAc WGA wheat germ ag.(Triticum vulgare) (GlcNAc-beta-)2-3; Sia