Glycolipids Isolated from the Spleen of Gaucher's Disease

Total Page:16

File Type:pdf, Size:1020Kb

Glycolipids Isolated from the Spleen of Gaucher's Disease Tohoku J. exp. Med., 1966, 88 , 277-388 Glycolipids Isolated from the Spleen of Gaucher's Disease Akira Makita, Chiyuki Suzuki and Zensaku Yosizawa Department of Biochemistry(Prof. Z. Yosizawa), Tohoku UniversitySchool of Medicine,Sendai and Tasuke Konno Department of Pediatrics (Prof. Ts. Arakawa), Tohoku UniversitySchool of Medicine,Sendai Gyyeolipids of Gaueher's spleens (three cases) were examined for their content and chemical characterization, comparing with normal control. Apparent accumulation of glucocerebroside was found in all cases. In a case, however, a faster-moving glycolipid fraction than cerebroside on a thin-layer chromatogram, traces of which were detected also in other cases and normal control, was found in a certain amount. From this fraction, a fatty acid ester of glucocerebroside was isolated. Two-thirds of fatty acid of this fraction were composed of the acids below C20. Hematoside which increased in a certain extent, had the same chemical structure, N-acetylneuraminoyl-(2•¨3)-galactosyl-(1•¨4)-glucosyl-ceramide, as that of normal human spleen. Other glycolipids separated from Gaucher's spleens showed no significant variation in amounts and were all identical in chemical character with those found already in normal spleen. Gaucher's disease, one of sphingolipidoses, has been well characterized by the accumulation of glycolipids in the reticuloendothelial system of various organs, especially of the spleen. In 1924, the deposited lipid in Gaucher's spleen was shown by Liebl to be cerebroside (cerasine). Since Aghion2 and Halliday et al.3 presented evidence that the accumulated cerebroside contains predominantly glucose, instead of galactose of cerebroside in central nervous system, many discussions4-7 have been focused on the constituent hexose of Gaucher's cerebroside. Recent investigations,8,9 however, established that the hexose moiety of cerebroside deposited in Gaucher's spleen was ex clusively glucose. In addition, the chemical structure10,11of Gaucher's cerebroside was confirmed to be identical with that of brain galactocerebroside except dif ferences of the species of its hexose and fatty acid. On the other hand, Makita and Yamakawa12 revealed that cerebroside isolat ed from non-Gaucher's spleen also contained predominantly glucose. Therefore, Received for publication, January 12, 1966. 277 278 A. Makita et al. they assumed that physiological glucocerebroside was accumulated abnormally in Gaucher's spleen. Recently, increased formation of other glycolipids in Gaucher's spleen as well as glucocerebroside was reported. That is, Philippart et al.13 showed the accumulation of hematoside-type mucolipid and ceramide dihexoside in addition to glucocerebroside in this disease. Brady et al.14 reported, moreover, the deficiency of an enzyme catalyzing conversion of glucocerebroside to ceramide in Gaucher's spleen. The present communication reports the results of the study on glycolipids of three cases of Gaucher's spleen, comparing with those of non-Gaucher spleens. In one case (acute infantile cerebral form) of these Gaucher's spleens, a cere broside-fatty acid ester and hematoside were found to be accumculated. Chemical structures of these compounds are proposed by the results of gas-chromatography on the methylated glycosides. MATERIALS AND METHODS The spleens were obtained surgically from 3 patients with Gaucher's disease. Case 1, an 1-year-old female with Gaucher's disease of acute cerebral form, kept frozen (128 g); Case 2, a 14-year-old male, of non-cerebral form, kept frozen (51 g,); and Case 3 , a 6-month-old male, of cerebral form, kept in formalin (11 g). As normal control, pooled seven spleens (82 g) from those who died at the age of 1-5 years were used. For quantitative isolation of glycolipids, the procedure of the preparation used earlier15 was partly modified by employing Folch's method.16 Typical preparation of glycolpids was carried out as follows: A portion (128 g) of the spleen in Case 1 was homogenized and extracted with 20 volumes (vow) of chloroform-methanol (2:1, v/v). The filtered extract was evaporated to dryness. The dried residue was extracted again with chloroform-methanol (2:1, v/v) to remove an insoluble material. This lipid mixture (6.4g, total lipids) was dissolved in hexane and then dialyzed against hexane through thin-rubber membrane.17 Outer hexane was changed twice. The non-dialyzable complex lipids were subjected to mild alkaline hydrolysis* in a mixture of each 100 ml of aqueous 0.6 N KOH and chloroform at 37•Ž for 1 hour under continuous stirring. After cooling and acidifying the reaction mixture to about pH 4 with 6 N HCl, each 100 ml of chloroform and methanol were added and shaken vigorously. When clear separation into two phases was not obtained, additional amount of methanol was introduced. By partition, the lower phase was separated and then dialyzed against water. The non-dialyzable fraction was freed from chloroform by * In Cases 2 and 3, a portion of the complex lipids, without alkaline treatment , was chromatographed directly on silicic acid by elution with 4% methanol in chloroform (96C-M), by which esterified glycolipids should be eluted. Glycolipids in Gaucher's Spleens 279 evaporation, lyophilyzed and dialyzed against hexane , as mentioned above, to remove free fatty acids . Furthermore, from the upper phase, small amount of glycolipids was obtained by the same method, and combined with the main crude sphingolipid fraction. A portion (1.1 g) of the substance (2.7 g, crude sphingolipid fraction) thus obtained was applied to a column of silicic acid (Fig. 1). Glycolipids in the eluates were checked by anthrone reaction as well as Fig. 1. Silicic acid column chromatogram of crude sphingolipids of Gau her's spleen (Case 1). Circles indicate the mobility of glycolipids on TLC developed with solvent I (chloroform-methanol-water, 65:25:4, v/v), detected by anthrone sulfuric reagent. Abbreviations: FG, faster glycolipid fraction; CMH, cerebroside; CDH, ceramide dihexoside; CTH, ceramide trihexoside; Glob, globoside; ML ‡U, hematoside. 90C-M indicates 90% chloroform in methanol. thin-layer chromatography (TLC) compared with the authentic compounds. By the first silicic acid chromatography, only sialic acid-containing glycolipid (ML ‡U) was obtained homogeneously as glycolipid on TLC (Fig. 1). This ML ‡U was further purified by using a column of diethylaminoethyl (DEAE-) cellulose18 to remove sphingomyelin. Other fractions containing two glycolipids were chromatographed successively on silicic acid or Florisil in a small scale, as described previously. 15 After one chromatographic treatment on silicic acid, 87 95% (w/w) of lipids was recovered. The amounts of the fractions (Frs. I-V of Fig. 1) obtained by the first silicic acid chromatography were estimated by weighing. As the glycolipid preparations were still contaminated with some phospholipids, the real amounts of the glycolipids were calculated by subtraction of the weight of phospholipids (P value•~25). Then, the amount of each 280 A. Makita et al . Fig. 2. Chromatograms of faster glycolipid fraction. Faster-glyco lipid fraction (95 mg) was applied to a column prepared from a slurry of silicic acid (8 g) and Hyflosupercel (4 g), eluting with 100 ml of each solvent (2-1). TLC (2-2) was developed by solvent ‡U . 1, Gaucher's cerebroside; 2, bovine brain cerebroside which was obtained from phospholipid containing cerebroside fraction by the alkaline treatment em ployed in the present study. Glycolipids in Gaucher's Spleens 281 glycolipid was determined from the proportion obtained by the final chromato graphic separation. Reprecipitation of lipid with acetone from the concentrated effiuents was avoided in all experiments described here, because a little loss of some glycolipids occurred by this procedure. Compositions of hexoses19 and fatty acids in glycolipids were determined by means of gas-chromatography (GLC). Based on the ratio of galactose to glucose thus obtained, the amount of hexose was estimated using anthrone and thionalide20 reagents. Other analytical methods15 and procedures of methyla tion followed by GLC of methylated glycosides21were described earlier, respectively. Chemical characterization of isolated compounds was carried out mainly in Case 1, and in the other cases the glycolipids separated were identified on TLC and by hexose compositions on GLC. RESULTS AND DISCUSSION 1. Cerebroside fatty acid ester. By the first chromatography (Fig. 1) of crude sphingolipids, a faster-moving glycolipid fraction (faster glycolipids) than cere broside on TLC appeared in Fr. I (Fig. 1) which contained cerebroside. This frac tion was subjected to Florisil chromatography by the elution with 96% chloroform in methanol (96C-M), 70C-M and 50C-M. The glycolipid fraction eluted with 96C-M was again dialyzed against hexane through rubber membrane. The non-dialyzable portion revealed to contain at least three glycolipids other than cerebroside by TLC in solvent ‡U (chloroform-methanol-water, 85:15:1.5, v/v). Therefore, it was rechromatographed on silicic acid (Fig. 2). A pure glycolipid (35 mg, Fr. ‡U of Fig. 2) was isolated by the elution with 98 C-M. This material showed an absorption at 1,730 cm-1 (ester carbonyl) in addition to those of cerebroside by infrared spectral analysis (Fig. 3). Chemical character: It was slightly yellow and partly soluble in acetone. Reactions of Liebermann-Burchard for cholesterol and Bial for sialic acid were negative. This compound had m.p. 104?-107?; [ƒ¿]18589-4.7? (c, 2.4; in pyridine); P, nil; ester value, 0.99 ,uequiv. /µmole (Snyder and Stephan's method22); hexose (as glucose), (16.9%) (thionalide reagent20). Glucose was comprised in 98% of total hexose and the remaining was galactose. Elemental analysis: C, 72.41; H, 11.95; N, 1.35. Diarachidyl-cerebroside, C64H123O9N (1049), requires C, 73.21; H, 11.73; N, 1.33; hexose, 17.2 On the bases of these data, this compound is most likely to be fatty acid ester of cerebroside. As the fatty acid composition in Table I indicates, the faster glycolipid fraction involved in this substance contained larger proportions of palmitic and stearic acids than those of cerebroside.
Recommended publications
  • Sphingolipid Metabolism in Cultured Fibroblasts: Microscopic And
    Proc. Nati Acad. Sci. USA Vol. 80, pp. 2608-2612, May 1983 Cell Biology Sphingolipid metabolism in cultured fibroblasts: Microscopic and biochemical studies employing a fluorescent ceramide analogue (Golgi apparatus/sphingomyelin/cerebrosides/liposomes/fluorescence) NAOMI G. LPSKY AND RICHARD E. PAGANO Department of Embryology, Carnegie Institution of Washington, 115 West University Parkway, Baltimore, Maryland 21210 Communicated by Harden M. McConnell, December 30, 1982 ABSTRACT A fluorescent analogue of ceramide, N-[7-(4-ni- HI trobenzo-2-oxa-1,3-diazole)]-e-aminocaproyl sphingosine (C6-NBD- ceramide), was used to investigate sphingolipid metabolism in HO-C-H Chinesehamster fibroblasts. C6-NBD-ceramide was incorporated | /~~~~0 into small unilamellar dioleoyl phosphatidylcholine vesicles and N N incubated with cells in monolayer culture at 20C, resulting in rapid 0 /9 and preferential transfer of the labeled ceramide from vesicles to HI ° cells. The cells were then washed and subsequently incubated at H-C-N- C-(CH2)5-N NO2 37°C for various intervals. The metabolism of C6-NBD-ceramide was monitored by lipid extraction and analysis, and the intracel- lular distribution of the labeled molecule was followed by fluo- H rescence microscopy. Initially, fluorescence was detected almost HO-C-C =C-(CH2)12- CH3 exclusively in mitochondria, with over 90% of the extractable lipid I fluorescence due to C6-NBD-ceramide. After 30 min at 370C, in- H H tense fluorescence. appeared in the Golgi apparatus. This organ- elle was identified by colocalization of NBD fluorescence with a FIG. 1. Structure of C6-NBD-ceramide. Golgi-apparatus-specific stain. At later times the plasma mem- brane became visibly labeled as well, at which point 90% of the orescent of the cell-associated fluorescence was recovered as NBD-labeled sphin- tracer allows direct microscopic observation gomyelin and NBD-labeled cerebroside.
    [Show full text]
  • Metabolism of Brain Glycolipid Fatty Acids '': Yasuo Kishimoto and Norman S
    Metabolism of Brain Glycolipid Fatty Acids '': Yasuo Kishimoto and Norman S. Radin, Mental Health Research Institute, University of Michigan, Ann Arbor, Michigan ABSTRACT and sulfatides contain NFA and tIFA, The metabolism of the fatty acid moieties saturated and unsaturated; the gangliosides, of brain cerebrosides, sulfatides, and however, contain only NFA in which there are gangliosides is reviewed and discussed. only traces of unsaturated acids. In the cere- The methodology involved in the isolation t)rosides and sulfatides there are two clusters of the fatty acids is described briefly. It of FA: those around 18 carbons long and those seems clear now that most of these acids around 24 carbons long. In the gangliosides are made by chain elongation of inter- there is only one cluster, centering around 18:0, mediate length fatty acids by addition of with negligible amounts of 22:0 and 24:0. acetate residues. The unsaturated acids Other points of contrast between gangliosides are made by desaturation of the inter- and the other two can be made: the former mediate length acids (palmitic, heptade- occurs primarily in brain gray matter, the canoic, stearic) followed by chain elonga- latter are primarily in white. The former tion. The hydroxy acids are made directly has glucose attached to the ceramide residue, from the corresponding nonhydroxy acids, the latter have galactose. The former has saturated, unsaturated, and odd-numbered. only traces of odd-numbered FA; the latter All the hydroxy acids undergo oxidative can contain considerable amounts of C~ and decarboxylation to yield fatty acids con- C2.~ FA. Further differences, particularly in taining one less carbon atom.
    [Show full text]
  • Sphingomyelin of Red Blood Cells in Lipidosis and in Dementia of Unknown Origin in Children
    Arch Dis Child: first published as 10.1136/adc.44.234.197 on 1 April 1969. Downloaded from Arch. Dis. Childh., 1969, 44, 197. Sphingomyelin of Red Blood Cells in Lipidosis and in Dementia of Unknown Origin in Children G. J. M. HOOGHWINKEL, H. H. VAN GELDEREN, and A. STAAL From the Laboratory of Medical Chemistry and the Departments of Paediatrics and Neurology, University of Leiden, The Netherlands Histological and chemical examinations of biopsy no diagnosis could be made, are briefly described in specimens from cerebral tissue of children suffering Table I. Incomplete investigation made it impossible to from undiagnosed progressive brain disease are reach a diagnosis in Cases 7 and 8. The molar con- performed increasingly. Important information centrations of phospholipids in the red blood cells therapeutic have been determined as described by Hooghwinkel is provided, which, though rarely of and Niekerk (1960), Hooghwinkel and Borri (1964), value, does increase precision of diagnosis, and Hooghwinkel, Borri, and Bruyn (1966). The prognosis, and genetic advice (Cumings, 1965a, amounts of the various phospholipids of red blood b, c; Poser, 1962; Adams, 1965). This is especially cells have been expressed as molar percentages of true of the chemical investigations, and it is likely total phospholipids. Absolute values of phospho- that chernical analyses will challenge the current lipids depend a good deal on size and shape of the classifications of progressive brain disease. Amaur- otic idiocy has already proved to be more hetero- 36 copyright. 0 geneous than was thought hitherto, while supposedly -a .A a different diseases, such as Niemann-Pick's and 0 34 .
    [Show full text]
  • Metabolism of Cerebroside Sulfate and Subcellular Distribution of Its
    Metabolism of Cerebroside Sulfate and Subcellular Distribution of Its Metabolites in Cultured Skin Fibroblasts from Controls, Metachromatic Leukodystrophy, and Globoid Cell Leukodystrophy Koji Inui, Masumi Furukawa, Shintaro Okada, and Hyakuji Yabuuchi Department ofPediatrics, Osaka University Medical School, Osaka 553, Japan Abstract each step result in metachromatic leukodystrophy (MLD), globoid cell leukodystrophy (GLD), and Farber disease. Due With pulse-chase study of 1-['4CJstearic acid-labeled cerebro- to enzyme deficiencies, massive lysosomal storage of lipids is side sulfate ('4C-CS) and subsequent subcellular fractionation demonstrated in MLD (4) and Farber disease (5). In GLD it is by Percoll gradient, the metabolism of CS and translocation of well known that there is no accumulation of galactosylcera- its metabolites in human skin fibroblasts from controls, meta- mide and that the major abnormalities are the presence of a chromatic leukodystrophy (MLD), and globoid cell leukodys- large number of globoid cells, a severe lack of myelin and trophy (GLD) were studied. In control skin fibroblasts, CS was astrogliosis in the nervous system (6). transported to lysosome and metabolized there to galactosyl- The enzymic defects of most lysosomal storage disorders ceramide (GalCer) and ceramide (Cer) within 1 h. During the have been clarified, but the molecular mechanisms that lead to chase period, radioactivity was increased at plasma membrane the clinical and pathological manifestations remain largely plus Golgi as phospholipids and no accumulation of GalCer or obscure. Recent morphological studies of neurons from Cer was found in lysosome. In MLD fibroblasts, 95% of '4C- humans (7), cats (8), and dogs (9) with gangliosidoses have CS taken up was unhydrolyzed at 24 h-chase and accumulated shown meganeurities, inappropriate proliferation ofsecondary at not only lysosome but also plasma membrane.
    [Show full text]
  • Properties and Units in the Clinical Laboratory Sciences Part X
    Pure Appl. Chem., Vol. 72, No. 5, pp. 747–972, 2000. © 2000 IUPAC INTERNATIONAL FEDERATION OF CLINICAL CHEMISTRY AND LABORATORY MEDICINE SCIENTIFIC DIVISION COMMITTEE ON NOMENCLATURE, PROPERTIES AND UNITS (C-NPU)# and INTERNATIONAL UNION OF PURE AND APPLIED CHEMISTRY CHEMISTRY AND HUMAN HEALTH DIVISION CLINICAL CHEMISTRY SECTION COMMISSION ON NOMENCLATURE, PROPERTIES AND UNITS (C-NPU)§ PROPERTIES AND UNITS IN THE CLINICAL LABORATORY SCIENCES PART X. PROPERTIES AND UNITS IN GENERAL CLINICAL CHEMISTRY (Technical Report) (IFCC–IUPAC 1999) Prepared for publication by HENRIK OLESEN1, INGE IBSEN1, IVAN BRUUNSHUUS1, DESMOND KENNY2, RENÉ DYBKÆR3, XAVIER FUENTES-ARDERIU4, GILBERT HILL5, PEDRO SOARES DE ARAUJO6, AND CLEM McDONALD7 1Office of Laboratory Informatics, Copenhagen University Hospital (Rigshospitalet), Copenhagen, Denmark; 2Dept. of Clinical Biochemistry, Our Lady’s Hospital for Sick Children, Dublin, Ireland; 3Dept. of Standardisation in Laboratory Medicine, Kommunehospitalet, Copenhagen, Denmark; 4Dept. of Clinical Biochemistry, Ciutat Sanitària i Universitària de Bellvitge, Barcelona, Spain; 5Dept. of Clinical Chemistry, Hospital for Sick Children, Toronto, Canada; 6Dept. of Biochemistry, IQUSP, São Paolo, Brazil; 7Regenstrief Inst. for Health Care, Indiana University School of Medicine, Indianapolis, Indiana, USA #§The combined Memberships of the Committee and the Commission (C-NPU) during the preparation of this report (1994 to 1996) were as follows: Chairman: H. Olesen (Denmark, 1989–1995); D. Kenny (Ireland, 1996). Members: X. Fuentes-Arderiu (Spain, 1991–1997); J. G. Hill (Canada; 1987–1997); D. Kenny (Ireland, 1994–1997); H. Olesen (Denmark, 1985–1995); P. L. Storring (UK, 1989–1995); P. Soares de Araujo (Brazil, 1994–1997); R. Dybkær (Denmark, 1996–1997); C. McDonald (USA, 1996–1997). Please forward comments to: H.
    [Show full text]
  • Metabolism of Cerebroside Sulfate and Subcellular Distribution of Its
    Metabolism of cerebroside sulfate and subcellular distribution of its metabolites in cultured skin fibroblasts from controls, metachromatic leukodystrophy, and globoid cell leukodystrophy. K Inui, … , S Okada, H Yabuuchi J Clin Invest. 1988;81(2):310-317. https://doi.org/10.1172/JCI113322. Research Article With pulse-chase study of 1-[14C]stearic acid-labeled cerebroside sulfate (14C-CS) and subsequent subcellular fractionation by Percoll gradient, the metabolism of CS and translocation of its metabolites in human skin fibroblasts from controls, metachromatic leukodystrophy (MLD), and globoid cell leukodystrophy (GLD) were studied. In control skin fibroblasts, CS was transported to lysosome and metabolized there to galactosylceramide (GalCer) and ceramide (Cer) within 1 h. During the chase period, radioactivity was increased at plasma membrane plus Golgi as phospholipids and no accumulation of GalCer or Cer was found in lysosome. In MLD fibroblasts, 95% of 14C-CS taken up was unhydrolyzed at 24 h-chase and accumulated at not only lysosome but also plasma membrane. In GLD fibroblasts, GalCer was accumulated throughout the subcellular fractions and more accumulated mainly at plasma membrane plus Golgi with longer pulse. This translocation of lipid from lysosome seems to have considerable function, even in lipidosis, which may result in an imbalance of the sphingolipid pattern on the cell surface and these changes might be one of causes of neuronal dysfunction in sphingolipidosis. Find the latest version: https://jci.me/113322/pdf Metabolism of Cerebroside Sulfate and Subcellular Distribution of Its Metabolites in Cultured Skin Fibroblasts from Controls, Metachromatic Leukodystrophy, and Globoid Cell Leukodystrophy Koji Inui, Masumi Furukawa, Shintaro Okada, and Hyakuji Yabuuchi Department ofPediatrics, Osaka University Medical School, Osaka 553, Japan Abstract each step result in metachromatic leukodystrophy (MLD), globoid cell leukodystrophy (GLD), and Farber disease.
    [Show full text]
  • Cerebroside Synthesis in Gaucher's Disease
    CEREBROSIDE SYNTHESIS IN GAUCHER'S DISEASE Eberhard G. Trams, Roscoe O. Brady J Clin Invest. 1960;39(10):1546-1550. https://doi.org/10.1172/JCI104175. Research Article Find the latest version: https://jci.me/104175/pdf CEREBROSIDE SYNTHESIS IN GAUCHER'S DISEASE* By EBERHARD G. TRAMS AND ROSCOE 0. BRADY (From the National Institute of Neurological Diseases and Blindness, Bethesda, Md.) (Submitted for publication April 12, 1960; accepted May 6, 1960) Familial lipodystrophic conditions, such as nectomized to arrest the pancytopenia of hyper- Gaucher's, Niemann-Pick, and Tay-Sachs dis- splenism. It was not possible to obtain control ease, are characterized by the intracellular ac- specimens of splenic tissue suitable for metabolic cumulation of abnormally large quantities of studies from normal children of the same age as sphingolipids. In Gaucher's disease, the offend- the patients with Gaucher's disease. Therefore, ing lipids are cerebrosides, while Niemann-Pick the results obtained in spleens from Gaucher's and Tay-Sachs diseases are characterized by the patients were compared with those obtained in accumulation of sphingomyelin and gangliosides, two children with Niemann-Pick disease and in respectively. In Gaucher's and Niemann-Pick one adult case of idiopathic thrombocytopenic disease, involvement of the reticuloendothelial sys- purpura. tem is extensive, and splenomegaly and hepato- Experiments were designed to explore several megaly are frequently observed. Lieb in 1924 factors that might be thought to contribute to the ( 1 ) identified the lipid stored in reticuloendothelial pathogenesis of Gaucher's disease. Enzymic dis- cells in a case of Gaucher's disease as a cerebro- turbances conceivably could lead to an alteration side.
    [Show full text]
  • Glucocerebrosidase: Functions in and Beyond the Lysosome
    Journal of Clinical Medicine Review Glucocerebrosidase: Functions in and Beyond the Lysosome Daphne E.C. Boer 1, Jeroen van Smeden 2,3, Joke A. Bouwstra 2 and Johannes M.F.G Aerts 1,* 1 Medical Biochemistry, Leiden Institute of Chemistry, Leiden University, Faculty of Science, 2333 CC Leiden, The Netherlands; [email protected] 2 Division of BioTherapeutics, Leiden Academic Centre for Drug Research, Leiden University, Faculty of Science, 2333 CC Leiden, The Netherlands; [email protected] (J.v.S.); [email protected] (J.A.B.) 3 Centre for Human Drug Research, 2333 CL Leiden, The Netherlands * Correspondence: [email protected] Received: 29 January 2020; Accepted: 4 March 2020; Published: 9 March 2020 Abstract: Glucocerebrosidase (GCase) is a retaining β-glucosidase with acid pH optimum metabolizing the glycosphingolipid glucosylceramide (GlcCer) to ceramide and glucose. Inherited deficiency of GCase causes the lysosomal storage disorder named Gaucher disease (GD). In GCase-deficient GD patients the accumulation of GlcCer in lysosomes of tissue macrophages is prominent. Based on the above, the key function of GCase as lysosomal hydrolase is well recognized, however it has become apparent that GCase fulfills in the human body at least one other key function beyond lysosomes. Crucially, GCase generates ceramides from GlcCer molecules in the outer part of the skin, a process essential for optimal skin barrier property and survival. This review covers the functions of GCase in and beyond lysosomes and also pays attention to the increasing insight in hitherto unexpected catalytic versatility of the enzyme. Keywords: glucocerebrosidase; lysosome; glucosylceramide; skin; Gaucher disease 1.
    [Show full text]
  • The Role of Sulfatide in Alzheimer's Disease
    Virginia Commonwealth University VCU Scholars Compass Theses and Dissertations Graduate School 2006 The Role of Sulfatide in Alzheimer's Disease Charles Britton Beasley Jr. Virginia Commonwealth University Follow this and additional works at: https://scholarscompass.vcu.edu/etd Part of the Nervous System Commons © The Author Downloaded from https://scholarscompass.vcu.edu/etd/1053 This Thesis is brought to you for free and open access by the Graduate School at VCU Scholars Compass. It has been accepted for inclusion in Theses and Dissertations by an authorized administrator of VCU Scholars Compass. For more information, please contact [email protected]. O Charles Britton Beasley, Jr., 2006 All Rights Reserved 'THE ROLE OF SULFATIDE IN ALZHEIMER'S DISEASE A thesis submitted in partial fulfillment of .the requirements for the degree of Master's in Anatomy and Neurobiology at Virginia Commonwealth University. by CHARLES BRITTON BEASLEY, JR. B.S. Biology Director: JEFFREY DUPREE, PHD ASSISTANT PROFESSOR, DEPARTMENT OF ANATOMY AND NEUROBIOLOGY Virginia Commonwealth University Richmond, Virginia August, 2006 Table of Contents Page List of Tables ................................................................................................................ iv List of Figures ............................................................................................................ v List of Abbreviations.................................................................................................. vi Chapter 1 Introduction .....................................................................................
    [Show full text]
  • Novel Lamprey Antibody Recognizes Terminal Sulfated Galactose Epitopes on Mammalian Glycoproteins
    ARTICLE https://doi.org/10.1038/s42003-021-02199-7 OPEN Novel lamprey antibody recognizes terminal sulfated galactose epitopes on mammalian glycoproteins Tanya R. McKitrick 1,8, Steffen M. Bernard2,8, Alexander J. Noll1,5,8, Bernard C. Collins2, Christoffer K. Goth 1,6, Alyssa M. McQuillan1, Jamie Heimburg-Molinaro1, Brantley R. Herrin3,7, ✉ Ian A. Wilson 2,4, Max D. Cooper 3 & Richard D. Cummings 1 The terminal galactose residues of N- and O-glycans in animal glycoproteins are often sia- lylated and/or fucosylated, but sulfation, such as 3-O-sulfated galactose (3-O-SGal), repre- 1234567890():,; sents an additional, but poorly understood modification. To this end, we have developed a novel sea lamprey variable lymphocyte receptor (VLR) termed O6 to explore 3-O-SGal expression. O6 was engineered as a recombinant murine IgG chimera and its specificity and affinity to the 3-O-SGal epitope was defined using a variety of approaches, including glycan and glycoprotein microarray analyses, isothermal calorimetry, ligand-bound crystal structure, FACS, and immunohistochemistry of human tissue macroarrays. 3-O-SGal is expressed on N- glycans of many plasma and tissue glycoproteins, but recognition by O6 is often masked by sialic acid and thus exposed by treatment with neuraminidase. O6 recognizes many human tissues, consistent with expression of the cognate sulfotransferases (GAL3ST-2 and GAL3ST- 3). The availability of O6 for exploring 3-O-SGal expression could lead to new biomarkers for disease and aid in understanding the functional roles of terminal modifications of glycans and relationships between terminal sulfation, sialylation and fucosylation. 1 Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA.
    [Show full text]
  • Labeling of Animal Cells with Fluorescent Dansyl Cerebroside Richard T
    Labeling of Animal Cells with Fluorescent Dansyl Cerebroside Richard T. C. Huang Institut für Virologie, Justus Liebig-Universität, Gießen (Z. Naturforsch. 31c, 737 — 740 [1976]; received September 9, 1976) Fluorescence Labeling, Dansyl Glycolipids, Glycolipid Patches, Plasma Membrane, Myxoviruses A dansyl (diaminonaphthalenesulfonyl)-derivative of cerebroside was prepared which could be effectively incorporated into the plasma membranes of tissue culture cells and erythrocytes. The cells which had assimilated the glycolipid fluoresced intensely and could be observed under a fluorescent microscope. Cells were initially labeled rather homogeneously over the whole surface. With longer incubation time organization of the fluorescent glycolipid took place and patches of the lipid in the membrane were formed. The redistribution and organization of the membrane lipid could be demonstrated most clearly when cells labeled with this fluorescent glycolipid were infected with myxoviruses. After infection of MDBK and BHK cells with fowl plague virus areas of dense fluorescence appeared at margines of neighboring cells. When BHK cells were infected with Newcastle disease virus fusion of the cells was accompanied by complete redistribution of the glycolipid. Erythrocytes could also easily incorporate dansyl cerebroside. Chicken erythrocytes which contain cytoplasmic and nuclear membranes incorporated the fluorescent glycolipid in both mem­ branes. TntvniliictioQ Material and Methods Fluorescent probes have been introduced into bio­ Preparation of dansyl cerebroside logical membranes in the past and changes in the intensity of fluorescence occurring have been mea­ Pure human brain cerebroside (5 g) was treated with barium hydroxide according to the procedure sured to interprete the fluidity and polarity of the of Klenk5 to yield psychosine (3.2 g).
    [Show full text]
  • Deficiency of the Hexosaminidase a Activator Protein in a Case of GM2 Gangliosidosis; Variant AB
    Pediatr. Res. 16: 2 17-222 (1982) Deficiency of the Hexosaminidase A Activator Protein in a Case of GM2 Gangliosidosis; Variant AB PETER HECHTMAN,'~"B. A. GORDON, AND N. M. K. NG YlNG KIN MRC Human Genetics Research Group and Centre for Human Genetics, Biology Department, McGill University, Montreal, Quebec, Canada [P. H.]; Children S Psychiatric Research Institute Biochemistry Laboratory, London, Ontario, Canada [B.A.G.]; and Donner Laboratory for Neurochemistry, Montreal Neurological Hospital, Montreal, Quebec, Canada [N.M. K. N. Y.K.] Summary Li and co-workers (20) and Hechtman (12) showed that the in vitro hydrolysis of the N-acetyl galactosaminyl linkage of GM2- A patient is described whose clinical course and pathologic ganglioside by purified preparations of human liver Hex A re- features, including massive brain storage of GM2 ganglioside in quires the presence of an activator (AP). Hydrolysis of synthetic grey matter, are identical with those of classical Tay-Sachs disease hexosaminidase substrates is unaffected by AP. This activator has despite normal levels of 8-N-acetyl hexosaminidase and normal been purified and shown to be a heat stable protein with a isozyme distribution. The kinetic properties and thermolability of molecular weight of 36,000 (13). An in vivo metabolic function for the patient's hexosaminidase are normal. Crude extracts of a the activator protein was demonstrated by Conzelman and postmortem sample of patient's liver can catalyze the hydrolysis Sandhoff (2) who showed that kidney extracts of the original AB of 5.1 pmoles of labeled GM2 ganglioside/l6 h/mg'of protein variant patient were deficient in the Hex A activator protein.
    [Show full text]