DOCSLIB.ORG

Sialic Acid Activation

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Sialic Acid Activation Glycobiology vol. 1 no. 5 pp. 441-447, 1991 MINI REVIEW Sialic acid activation Edward L.Kean a few instances has the sugar nucleotide actually been measured in animal tissues. Harms et al. (1973) determined the con- Department of Ophthalmology and the Department of Biochemistry, Case Western Reserve University, Cleveland, OH 44106, USA centration of CMP-NeuAc in rat liver to be 40.8 nmol/g tissue. The pool size of CMP-NeuAc in Chinese hamster ovary cells Cytidine 5-monophosphosialic acid (CMP-sialic acid) is the was measured by Briles et al. (1977) (1.6 nmol/mg cell activated form of sialic acid which is required for the bio- protein). Corfield et al. (1976) reported the presence of CMP- synthesis of sialic acid-containing complex carbohydrates. NeuAc and CMP-9-O-Ac-NeuAc in bovine submandibular Its discovery over 30 years ago by the laboratory of Dr Saul glands, and provided a partial identification. Carey and Downloaded from https://academic.oup.com/glycob/article/1/5/441/621435 by guest on 01 October 2021 Roseman was a landmark in research dealing with the bio- Hirschberg (1979) isolated CMP-NeuAc from mouse liver and synthesis of these compounds. A review is presented of the determined its concentration (37 nmol/g). salient features concerning this molecule: its discovery, The enzymatic synthesis of CMP-NeuAc was reported by chemistry, biosynthesis, subcellular location, enzymatic Roseman (1962) using a preparation from hog submaxillary cleavage, transport and molecular biology. This report does glands, and by Warren and Blacklow (1962a, b) using a pre- not deal with its utilization by the sialyltransferases. paration from Neisseria meningitidis. CMP-sialic acid is synthesized by the following reaction: CTP + sialic acid -• Key words: cytidine 5'-monophosphosialic acid/CMP-sialic CMP-sialic acid + PPi. Unlike the synthesis of other sugar acid/synthetase/hydrolase/subcellular location nucleotides which involves a reaction between a nucleotide triphosphate and a sugar-1-phosphate, the biosynthesis of CMP- sialic acid is unique in using the non-phosphorylated sugar donor (a reaction which also occurs in the formation of CMP- KDO). Subsequently, several laboratories described the syn- Properties of CMP-NeuAc and CMP-NeuAc synthetase thesis of CMP-NeuAc, among which are reports by Shoyab et al. (1964) using a preparation from sheep brain; Kean and In 1959, Comb, Shimizu and Roseman reported the discovery Roseman (1966a, b) using preparations from hog submaxillary of a new sugar nucleotide, CMP-N-acetylneuraminic acid gland and E.coli; van den Eijnden et al. (1972) with a calf brain (CMP-NeuAc)1. The compound was isolated from the cells of preparation; Hultsch et al. (1972), from rat liver; Schauer Escherichia coli K-235, the organism that produces a homo- et al. (1980) on a frog liver preparation. The properties of the polymer of sialic acid, colominic acid. Now, some 30 years compounds synthesized by these sources were similar in terms later, the gene governing its synthesis has been cloned. The of composition, chemical and enzymatic reactivity and stability, intervening years saw the wide demonstration that CMP-sialic and chromatographic properties such as great sensitivity to acid was the activated form of sialic acid required for the bio- acid; relatively stable at neutral or alkaline pH; resistance to the synthesis of sialic acid-containing complex carbohydrates. The actions of 5' nucleotidase, phosphodiesterase, phosphatase, discovery was a key one in furthering our understanding of the neuraminaldolase and neuraminidases; under acid conditions biosynthesis of this class of compounds. Although but a one- they underwent typical colorimetric reactions for sialic acid; page letter to the Editor, this initial report revealed many of the resistance to reduction with NaBHj. In general, the enzymes properties of this unique compound. This preliminary com- synthesizing CMP-NeuAc from all sources examined also had munication was followed by an extensive characterization of similar properties, as summarized in Table I. Unlike the CMP-NeuAc from E.coli K-235 (Comb et al., 1966). The most enzymes from N.meningitidis and E.coli K-235, the prepara- unique characteristic of the compound was the presence of one tions from the animal tissues (submaxillary gland, sheep brain, instead of two phosphate residues as is found in other sugar retina, rat liver) also functioned with N-glycolylneuraminic nucleotides, a property shared with only one other molecule of acid (NeuGc) as substrate. Also, unlike the enzyme from this type, CMP-3-deoxy-D-manno-octulosonate (CMP-KDO; N. meningitidis, the reaction carried out by the hog submaxillary Ghalambor and Heath, 1966), a compound involved in bacterial enzyme was shown to be reversible (Kean and Roseman, cell wall biosynthesis. 1966a, b); the ratio of the forward reaction to the reverse While the biosynthesis of CMP-NeuAc has been readily reaction was estimated to be - 1000:1. Reversibility could not demonstrated in a variety of animal tissues (see below), in only be detected with the enzyme from sheep brain (Shoyab et al., 1964). (This property was not reported in the other tissues.) The possibility that carboxyl activation might be involved in the 'The nomenclature in common use in this field is followed in which the term 'sialic acid' is the family name of the group of nine carbon amino sugar formation of CMP-NeuAc by the mammalian enzyme was acids having an acyl group on the amino nitrogen. Individual sialic acids ruled out in view of the lack of hydroxamate formation during are derivatives of neuraminic acid, such as AZ-acetylneuraminic acid or NeuAc, the reaction (Kean and Roseman, 1966a). whose systematic name is 5-acetamido-3,5-dideoxy-D-glycero-r>-galacto- The anomeric configuration about carbon atom 2 of the sialic nonulosonic acid. acid portion of CMP-NeuAc has been controversial. On the This review is dedicated to Dr Saul Roseman as part of a symposium in his honour at the 1 lth International Symposium on Glycoconjugates, held on June basis of its chemical properties and optical rotation, and 30-July 5, 1991, in Toronto, Canada. assuming that the Hudson isorotation rules applied, the linkage © Oxford University Press 441 E.L.Kean TaMe I. Comparison of properties of CMP-sialic acid synthetases Warren and Blacklow Kean and Roseman Shoyab van den Eijnden Kean Van Dijk Schauer (1962a, b) (1966a, b) et al. (1964) et al. (1972) (1970) et al. (1973) et al. (1980) Form of enzyme Soluble Soluble Soluble Soluble Nuclear Nuclear Soluble Tissue Neisseria meningitidis Hog submaxillary Sheep brain Calf brain Hog retina, Kidney cortex Frog liver rat liver Km (NeuAc) 0.25 mM 0.8 mM 06 mM 0 90 mM 0.72 mM 1.6 mM 1.6 mM Km (NeuGc) Inactive 2.3 mM 2.3 mM - 1.40 mM - 2 3 mM Km (CTP) 0.56 mM 060 mM 1.8 mM 0.50 mM 0.48 mM 1.6 mM 0 6 mM pH optimum 8.5 90 8.0 9.0 8.5 8.5-9.0 9.0 Fold purified 205^03 264-528 39 4.9 - - 250 Sulphydryl requirement Required Stabilizing Stimulatory Present Stabilizing Stabilizing Present Downloaded from https://academic.oup.com/glycob/article/1/5/441/621435 by guest on 01 October 2021 Reversibility No Yes No - - - - Stability Freeze/thaw Crude, stable Crude, stable - Stable Unstable Unstable Purified, unstable Purified, unstable Purified, stable 4°C Stable (2 weeks) Stable (3 weeks) Stable Stable Stable (1 week) Stable, 1 week (2 weeks) Metal; optimal Mg; (25 mM) Mg; (20 mM) Mg; (30 mM) Mg; (40 mM) Mg; (20 mM) Mg; (3.0 mM) Mg or Ca; concentration (50 mM) was suggested to be (3 by Comb et al. (1966). This assignment 1971). The nuclear location of the sialic acid-activating enzyme received support by the study of Haverkamp et al. (1979) who was firmly established by studies using highly purified nuclei examined the anomeric configuration by means of I4C NMR from hog retina, rat liver, kidney, spleen and brain (Kean, spectroscopy. Stone and Kolodny (1971), however, have con- 1970, 1972a). When corrected for the yield of nuclei, from cluded from circular dichroism studies of CMP-NeuAc that the 54 to 90% of the total activity could be ascribed to the nuclei ketosidic linkage is a. Indirect support for the /3 configuration from these several tissues. The kinetic properties and stability may be obtained from the investigations of Yu and Ledeen of the nuclear enzyme were similar to those of the soluble form (1969), who examined the stereochemistry of the two anomeric described previously, as seen in Table I. methyl ketosides of NeuAc. The configuration to which the With the possibility that the nuclear location was due to a-D assignment was made was susceptible to the action of artifactual redistribution of cellular constituents as a result of neuraminidase, while the /3 ketoside was resistant. CMP-NeuAc homogenization, the nuclear location of CMP-NeuAc syn- has been observed to be resistant to the action of neuraminidase, thetase was examined further by employing a procedure to consistent with the /3 assignment. Since the configuration of fractionate the cell that did not involve homogenization. Harvey sialic acid in glycoconjugates occurs only with the a con- (1956) demonstrated that unfertilized sea urchin eggs could be figuration, a single inversion of configuration would, therefore, segmented into nucleate and non-nucleate portions merely by accompany the transfer of sialic acid from the sugar nucleotide centrifugation. Fractions (halves and quarters of the eggs) thus to the acceptor. enriched with nuclei can be obtained by this mild procedure. The distribution of CMP-sialic acid synthetase was followed in the nucleate and anucleate segments, and was observed to accompany the nuclear-enriched fractions (Kean and Bruner, Subcellular location; nuclear CMP-NeuAc synthetase 1971), an observation consistent with the nuclear location of the CMP-NeuAc synthetase examined in each of the reports enzyme.
Load more

Recommended publications
  • Terminal Sialic Acid Linkages Determine Different Cell Infectivities of Human Parainfluenza Virus Type 1 and Type 3
    Virology 464-465 (2014) 424–431 Contents lists available at ScienceDirect Virology journal homepage: www.elsevier.com/locate/yviro Terminal sialic acid linkages determine different cell infectivities of human parainfluenza virus type 1 and type 3 Keijo Fukushima a,1, Tadanobu Takahashi a,1, Seigo Ito a, Masahiro Takaguchi a, Maiko Takano a, Yuuki Kurebayashi a, Kenta Oishi a, Akira Minami a, Tatsuya Kato b,f, Enoch Y Park b,e,f, Hidekazu Nishimura c, Toru Takimoto d, Takashi Suzuki a,n a Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka 4228526, Japan b Laboratory of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 4228529, Japan c Virus Research Center, Sendai Medical Center, 2-8-8 Miyagino, Miyagino-ku, Sendai, Miyagi 9838520, Japan d Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, NY 14642, USA e Laboratory of Biotechnology, Integrated Bioscience Section, Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 4228529, Japan f Laboratory of Biotechnology, Green Chemistry Research Division, Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 4228529, Japan article info abstract Article history: Human parainfluenza virus type 1 (hPIV1) and type 3 (hPIV3) initiate infection by sialic acid binding. Received 22 May 2014 Here, we investigated sialic acid linkage specificities for binding and infection of hPIV1 and hPIV3 by Returned to author for revisions using sialic acid linkage-modified cells treated with sialidases or sialyltransferases. The hPIV1 is bound to 8 July 2014 only α2,3-linked sialic acid residues, whereas hPIV3 is bound to α2,6-linked sialic acid residues in Accepted 11 July 2014 addition to α2,3-linked sialic acid residues in human red blood cells.
    [Show full text]
  • Sialic Acids and Their Influence on Human NK Cell Function
    cells Review Sialic Acids and Their Influence on Human NK Cell Function Philip Rosenstock * and Thomas Kaufmann Institute for Physiological Chemistry, Martin-Luther-University Halle-Wittenberg, Hollystr. 1, D-06114 Halle/Saale, Germany; [email protected] * Correspondence: [email protected] Abstract: Sialic acids are sugars with a nine-carbon backbone, present on the surface of all cells in humans, including immune cells and their target cells, with various functions. Natural Killer (NK) cells are cells of the innate immune system, capable of killing virus-infected and tumor cells. Sialic acids can influence the interaction of NK cells with potential targets in several ways. Different NK cell receptors can bind sialic acids, leading to NK cell inhibition or activation. Moreover, NK cells have sialic acids on their surface, which can regulate receptor abundance and activity. This review is focused on how sialic acids on NK cells and their target cells are involved in NK cell function. Keywords: sialic acids; sialylation; NK cells; Siglecs; NCAM; CD56; sialyltransferases; NKp44; Nkp46; NKG2D 1. Introduction 1.1. Sialic Acids N-Acetylneuraminic acid (Neu5Ac) is the most common sialic acid in the human organism and also the precursor for all other sialic acid derivatives. The biosynthesis of Neu5Ac begins in the cytosol with uridine diphosphate-N-acetylglucosamine (UDP- Citation: Rosenstock, P.; Kaufmann, GlcNAc) as its starting component [1]. It is important to understand that sialic acid T. Sialic Acids and Their Influence on formation is strongly linked to glycolysis, since it results in the production of fructose-6- Human NK Cell Function. Cells 2021, phosphate (F6P) and phosphoenolpyruvate (PEP).
    [Show full text]
  • Review Sialic Acid-Specific Lectins: Occurrence, Specificity and Function
    Cell. Mol. Life Sci. 63 (2006) 1331–1354 1420-682X/06/121331-24 DOI 10.1007/s00018-005-5589-y Cellular and Molecular Life Sciences © Birkhäuser Verlag, Basel, 2006 Review Sialic acid-specific lectins: occurrence, specificity and function F. Lehmanna, *, E. Tiralongob and J. Tiralongoa a Institute for Glycomics, Griffith University (Gold Coast Campus), PMB 50 Gold Coast Mail Centre Australia 9726 (Australia), Fax: +61 7 5552 8098; e-mail: [email protected] b School of Pharmacy, Griffith University (Gold Coast Campus), PMB 50 Gold Coast Mail Centre Australia 9726 (Australia) Received 13 December 2005; received after revision 9 February 2006; accepted 15 February 2006 Online First 5 April 2006 Abstract. Sialic acids consist of a family of acidic nine- through specific interactions with lectins, a family of carbon sugars that are typically located at the terminal po- proteins that recognise and bind sugars. This review will sitions of a variety of glycoconjugates. Naturally occur- present a detailed overview of our current knowledge re- ring sialic acids show an immense diversity of structure, garding the occurrence, specificity and function of sialic and this reflects their involvement in a variety of biologi- acid-specific lectins, particularly those that occur in vi- cally important processes. One such process involves the ruses, bacteria and non-vertebrate eukaryotes. direct participation of sialic acids in recognition events Keywords. Sialic acid, lectin, sialoglycoconjugate, sialic acid-specific lectin, adhesin, infectious disease, immunology. Introduction [1, 2]. The largest structural variations of naturally occurring Sia are at carbon 5, which can be substituted with either an Sialic acids (Sia) are a family of nine-carbon a-keto acids acetamido, hydroxyacetamido or hydroxyl moiety to form (Fig.
    [Show full text]
  • Direct Determination of Sialic Acids in Glycoprotein Hydrolyzates by HPAE-PAD
    Application Update 180 Update Application Direct Determination of Sialic Acids in Glycoprotein Hydrolyzates by HPAE-PAD Thermo Fisher Scientific, Inc. INTRODUCTION In this work, sialic acids are determined in five Sialic acids are critical in determining glycoprotein representative glycoproteins by acid hydrolysis followed bioavailability, function, stability, and metabolism.1 by HPAE-PAD. Sialic acid determination by HPAE-PAD Although over 50 natural sialic acids have been identified,2 on a Thermo Scientific™ Dionex™ CarboPac™ PA20 two forms are commonly determined in glycoprotein column is specific and direct, eliminating the need for products: N-acetylneuraminic acid (Neu5Ac) and sample derivatization after sample preparation. The use N-glycolylneuraminic acid (Neu5Gc). Because humans of a disposable gold on polytetrafluoroethylene (Au on do not generally produce Neu5Gc and have been shown PTFE) working electrode simplifies system maintenance to possess antibodies against Neu5Gc, the presence of this compared to conventional gold electrodes while providing sialic acid in a therapeutic agent can potentially lead to an consistent response with a four-week lifetime. The rapid immune response.3 Consequently, glycoprotein sialylation, gradient method discussed separates Neu5Ac and Neu5Gc and the identity of the sialic acids, play important roles in under 10 min with a total analysis time of 16.5 min, in therapeutic protein efficacy, pharmacokinetics, and compared to 27 min using the Dionex CarboPac PA10 potential immunogenicity. column by a previously published method.6,7 By using the Dionex CarboPac PA20 column, the total analysis time is Sialic acid determination can be performed by many reduced, eluent consumption and waste generation are methods. Typically, sialic acids are released from glyco- reduced, and sample throughput is improved.
    [Show full text]
  • REVIEW the Role and Potential of Sialic Acid in Human Nutrition
    European Journal of Clinical Nutrition (2003) 57, 1351–1369 & 2003 Nature Publishing Group All rights reserved 0954-3007/03 $25.00 www.nature.com/ejcn REVIEW The role and potential of sialic acid in human nutrition B Wang1* and J Brand-Miller1 1Human Nutrition Unit, School of Molecular and Microbial Biosciences, University of Sydney, NSW, Australia Sialic acids are a family of nine-carbon acidic monosaccharides that occur naturally at the end of sugar chains attached to the surfaces of cells and soluble proteins. In the human body, the highest concentration of sialic acid (as N-acetylneuraminic acid) occurs in the brain where it participates as an integral part of ganglioside structure in synaptogenesis and neural transmission. Human milk also contains a high concentration of sialic acid attached to the terminal end of free oligosaccharides, but its metabolic fate and biological role are currently unknown. An important question is whether the sialic acid in human milk is a conditional nutrient and confers developmental advantages on breast-fed infants compared to those fed infant formula. In this review, we critically discuss the current state of knowledge of the biology and role of sialic acid in human milk and nervous tissue, and the link between sialic acid, breastfeeding and learning behaviour. European Journal of Clinical Nutrition (2003) 57, 1351–1369. doi:10.1038/sj.ejcn.1601704 Keywords: sialic acid; ganglioside; sialyl-oligosaccharides; human milk; infant formula; breastfeeding Introduction promising new candidate is sialic acid (also known as The rapid growth and development of the newborn infant N-acetylneuraminic acid), a nine-carbon sugar that is a puts exceptional demands on the supply of nutrients.
    [Show full text]
  • A Structure-Based Rationale for Sialic Acid Independent Host-Cell Entry of Sosuga Virus
    A structure-based rationale for sialic acid independent host-cell entry of Sosuga virus Alice J. Stelfoxa and Thomas A. Bowdena,1 aDivision of Structural Biology, Wellcome Centre for Human Genetics, University of Oxford, OX3 7BN Oxford, United Kingdom Edited by Peter Palese, Icahn School of Medicine at Mount Sinai, New York, NY, and approved September 9, 2019 (received for review April 23, 2019) The bat-borne paramyxovirus, Sosuga virus (SosV), is one of many myxoviral RBPs consist of an N-terminal cytoplasmic region, paramyxoviruses recently identified and classified within the transmembrane domain, stalk region, and C-terminal six-bladed newly established genus Pararubulavirus, family Paramyxoviridae. β-propeller receptor-binding domain. Paramyxoviral RBPs The envelope surface of SosV presents a receptor-binding protein organize as dimer-of-dimers on the viral envelope, with the (RBP), SosV-RBP, which facilitates host-cell attachment and entry. receptor-binding heads forming dimers and the stalk regions driving Unlike closely related hemagglutinin neuraminidase RBPs from tetramization through disulphide bonding (18–24). Paramyxoviral other genera of the Paramyxoviridae, SosV-RBP and other para- RBPs functionally categorize into three groups: hemagglutinin- rubulavirus RBPs lack many of the stringently conserved residues neuraminidase (HN), hemagglutinin (H), and glycoprotein (G) required for sialic acid recognition and hydrolysis. We determined (25). Unlike HN RBPs, which recognize and hydrolyze sialic the crystal structure of the globular head region of SosV-RBP, acid presented on host cells, H and G RBPs attach to proteinous revealing that while the glycoprotein presents a classical para- receptors, such as SLAMF1 (26–28) and ephrin receptors (29, myxoviral six-bladed β-propeller fold and structurally classifies 30), respectively.
    [Show full text]
  • Defining Virus–Sialic Acid Interactions
    REVIEWS The sweet spot: defining virus–sialic acid interactions Jennifer E. Stencel-Baerenwald1,2*, Kerstin Reiss3,4*, Dirk M. Reiter3,5, Thilo Stehle3,6 and Terence S. Dermody1,2,6 Abstract | Viral infections are initiated by attachment of the virus to host cell surface receptors, including sialic acid-containing glycans. It is now possible to rapidly identify specific glycan receptors using glycan array screening, to define atomic-level structures of virus–glycan complexes and to alter the glycan-binding site to determine the function of glycan engagement in viral disease. This Review highlights general principles of virus–glycan interactions and provides specific examples of sialic acid binding by viruses with stalk-like attachment proteins, including influenza virus, reovirus, adenovirus and rotavirus. Understanding virus–glycan interactions is essential to combating viral infections and designing improved viral vectors for therapeutic applications. Viral attachment to receptors that are expressed on host has yielded general principles of virus–glycan interac- cells initiates infection and therefore, viral receptors are tions that may aid in the design of antiviral drugs and determinants of host range and govern host cell suscep- viral vectors. 1Department of Pathology, tibility. Various cell surface carbohydrates, including Microbiology, and sialylated glycans1–6, glycosaminoglycans7–10 and human Virus–sialic acid interactions Immunology, Vanderbilt 11,12 University School of Medicine. blood group antigens (HBGAs) , function as host cell Sialic acids are derivatives of neuraminic acid, which 2Elizabeth B. Lamb Center receptors for viral attachment and entry. is a nine-carbon monosaccharide that is ubiquitously for Pediatric Research, Although viruses have been known for some time expressed in higher vertebrates15.
    [Show full text]
  • Influenza Neuraminidase Operates Via a Nucleophilic Mechanism and Can
    ARTICLE Received 27 Sep 2012 | Accepted 15 Jan 2013 | Published 19 Feb 2013 DOI: 10.1038/ncomms2487 Influenza neuraminidase operates via a nucleophilic mechanism and can be targeted by covalent inhibitors Christopher J. Vavricka1,2,*, Yue Liu2,*, Hiromasa Kiyota3, Nongluk Sriwilaijaroen4,5, Jianxun Qi2, Kosuke Tanaka3, Yan Wu 2, Qing Li2,6, Yan Li2, Jinghua Yan2, Yasuo Suzuki5 & George F. Gao1,2,6,7 Development of novel influenza neuraminidase inhibitors is critical for preparedness against influenza outbreaks. Knowledge of the neuraminidase enzymatic mechanism and transition- state analogue, 2-deoxy-2,3-didehydro-N-acetylneuraminic acid, contributed to the devel- opment of the first generation anti-neuraminidase drugs, zanamivir and oseltamivir. However, lack of evidence regarding influenza neuraminidase key catalytic residues has limited strategies for novel neuraminidase inhibitor design. Here, we confirm that influenza neuraminidase conserved Tyr406 is the key catalytic residue that may function as a nucleophile; thus, mechanism-based covalent inhibition of influenza neuraminidase was conceived. Crystallographic studies reveal that 2a,3ax-difluoro-N-acetylneuraminic acid forms a covalent bond with influenza neuraminidase Tyr406 and the compound was found to possess potent anti-influenza activity against both influenza A and B viruses. Our results address many unanswered questions about the influenza neuraminidase catalytic mechanism and demonstrate that covalent inhibition of influenza neuraminidase is a promising and novel strategy for the development of next-generation influenza drugs. 1 Research Network of Immunity and Health (RNIH), Beijing Institutes of Life Science (BIOLS), Beijing 100101, China. 2 CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
    [Show full text]
  • Sialic Acid Content of the Erythrocyte and of an Ascites Tumor Cell of the Mouse
    Sialic Acid Content of the Erythrocyte and of an Ascites Tumor Cell of the Mouse AARON MILLER, JQHN F. SULLIVAN, AND JAY H. KATZ (P.AJ4ioisotope and Medical Services of the Boston Veterans Adminjairation Hospital, and the Department of Medicine, Boston University @SchoolofMedicine, Boston, Massachusetts) SUMMARY The sialic acid content of the mouse erythrocyte was compared with that of the Ehrlich ascites carcinoma cell. Neuraminidase treatment of intact tumor cells yielded 36 times more sialic acid than was released from the erythrocyte on a per cell basis. If this were all located on the surface, it may be calculated that the density of sialic acid on the Ehrlich ascites carcinoma cell is about 4 times greater than that on the erythro cyte. Trypsin released a sialic acid-containing fragment from both types of cells, the sialic acid being in the “bound―form.The sialic acid released by trypsin treatment of Ehrlich ascites tumor cells was only about one-sixth of that released by neuraminidase. By contrast, approximately equal amounts were liberated from the erythrocytes by these two enzymes. The erythrocyte sialic acid-containing substance was nondialyzable and was not precipitated by trichioroacetic acid. N-acetylneuraminic acid constituted approximately 70 per cent and N-glycolyneuraminic acid the remaining 30 per cent of the sialic acid of the tumor cell as determined by paper chromatography. In the erythrocyte N-acetylneuraminic acid constituted almost all the sialic acid, N-glycolyl neuraminic acid being undetectable in this cell. The sialic acids are a family of substances ured isoelectric point of about pH 4.0 (18).
    [Show full text]
  • Evolution and Diversity of Bat and Rodent Paramyxoviruses from North America Brendan B
    bioRxiv preprint doi: https://doi.org/10.1101/2021.07.01.450817; this version posted July 2, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Evolution and diversity of bat and rodent Paramyxoviruses from North America Brendan B. Larsen1, Sophie Gryseels1,2*, Hans W. Otto1, Michael Worobey1 1Department of Ecology and Evolutionary Biology, University of Arizona, Tucson, AZ 2 Department of Microbiology, Immunology and Transplantation, Rega Institute, KU Leuven, Laboratory of Clinical and Evolutionary Virology, Leuven, Belgium * Current addresses: Evolutionary Ecology group, Department Biology, University of Antwerp, Belgium OD Taxonomy and Phylogeny, Royal Belgian Institute of Natural Sciences, Belgium Abstract Paramyxoviruses are a diverse group of negative-sense, single-stranded RNA viruses of which several species cause significant mortality and morbidity. In recent years the collection of paramyxoviruses sequences detected in wild mammals has substantially grown, however little is known about paramyxovirus diversity in North American mammals. To better understand natural paramyxovirus diversity, host range, and host specificity, we sought to comprehensively characterize paramyxoviruses across a range of diverse co-occurring wild small mammals in Southern Arizona. We used highly degenerate primers to screen fecal and urine samples and obtained a total of 55 paramyxovirus sequences from 12 rodent species and 6 bat species. We also performed illumina RNA-seq and de novo assembly on 14 of the positive samples to recover a total of 5 near full-length viral genomes.
    [Show full text]
  • Sialylation: a Critical Glycosylation Quality Attribute for Biopharmaceuticals J.L
    Sialylation: A Critical Glycosylation Quality Attribute for Biopharmaceuticals J.L. Hendel, R.P. Kozak, C.L. Morgan, L. Royle Ludger Ltd., Culham Science Centre, Oxfordshire, OX14 3EB, United Kingdom [email protected] Introduction Qualitative Standards: Sialic Acid reference Panel (SRP) and Neu5,9Ac2 Biopharmaceutical Sialic Acid Analysis to Satisfy Regulatory Over the last two years the vast majority (27 of 28) of the FDA approved therapeutic biologics have been A sialic acid reference panel (SRP) containing a mixture of sialic acids found in humans and animals is used as a system suitability standard. The sialic Requirements glycoprotiens (1). These glycosylated therapeutics are comprised of glycoprotein hormones, cytokines, clotting acid reference standard contains Neu5Gc, Neu5Ac, Neu5,7Ac2, Neu5Gc9Ac, Neu5,9Ac2, and Neu5,7,(8),9Ac3. factors and monoclonal antibodies. Sialic acids are terminal, negatively charged monosaccharides present on Sialic acid analysis is a regulatory requirement and should be performed throughout the product lifecycle. The key many N- and O-glycans. Biopharmaceuticals often contain two main types of sialic acid; N-acetyl-neuraminic acid The acceptance criteria for this standard are that the HPLC profiles from the SRP at the start and end of the sample set should overlap with minimal data acquired from this study: (Neu5Ac, purple box figure 1) and N-glycolyl-neuraminic acid (Neu5Gc, blue box figure 1). Neu5Ac is found in both drift in retention time (e.g. ± 0.1 min.). The DMB profile for the SRP is shown in Figure 2A. 1. Identity of each sialic acid present human and non-human cells, whereas Neu5Gc not present on human glycoproteins and is immunogenic (2).
    [Show full text]
  • Decrease of Sialic Acid Residues As an Eat-Me Signal on the Surface of Apoptotic Lymphocytes
    Research Article 3347 Decrease of sialic acid residues as an eat-me signal on the surface of apoptotic lymphocytes Hanna Marie Meesmann1, Eva-Marie Fehr1, Sonja Kierschke1, Martin Herrmann2, Rostyslav Bilyy3, Petra Heyder1, Norbert Blank1, Stefan Krienke1, Hanns-Martin Lorenz1 and Martin Schiller1,* 1Department of Medicine V, University of Heidelberg, Im Neuenheimer Feld 410, 69120 Heidelberg, Germany 2Institute for Clinical Immunology and Rheumatology, Department of Internal Medicine 3, University Hospital Erlangen, 91054 Erlangen, Germany 3Institute of Cell Biology, National Academy of Sciences of Ukraine, Drahomanov Street 14/16, 79005 Lviv, Ukraine *Author for correspondence ([email protected]) Accepted 14 June 2010 Journal of Cell Science 123, 3347-3356 © 2010. Published by The Company of Biologists Ltd doi:10.1242/jcs.066696 Summary The silent clearance of apoptotic cells is essential for cellular homeostasis in multicellular organisms, and several mediators of apoptotic cell recognition have been identified. However, the distinct mechanisms involved are not fully deciphered yet. We analyzed alterations of the glycocalyx on the surfaces of apoptotic cells and its impact for engulfment. After apoptosis induction of lymphocytes, a decrease of 2,6-terminal sialic acids and sialic acids in 2,3-linkage with galactose was observed. Similar changes were to be found on the surface of apoptotic membrane blebs released during early stages of apoptosis, whereas later released blebs showed no impaired, but rather an increased, exposure of sialic acids. We detected an exposure of fucose residues on the surface of apoptotic-cell-derived membrane blebs. Cleavage by neuraminidase of sialic acids, as well as lectin binding to sialic acids on the surfaces, enhanced the engulfment of apoptotic cells and blebs.
    [Show full text]