Cara-Lynne Schengrund

Biological roles of sialic acid

1 Chemistry and Analysis of Sialic Acid.- I. Historical Background.- II. Natural Occurrence of Sialic Acids.- III. Isolation and Purification.- IV. Chemistry of Sialic Acids.- A. Basic Structures.- B. Stereochemistry.- C. Chemical Reactions and Derivatives.- V. Synthesis.- VI. Quantification of Sialic Acids.- A. Colorimetric and Fluorometric Assays.- B. Enzymatic Assay.- C. Gas-Liquid Chromatography.- VII. References.- 2 The Natural Occurrence of Sialic Acids.- I. Introduction.- II. The Natural Occurrence of Sialic Acids.- A. Viruses.- B. Bacteria.- C. Plants.- D. Invertebrates.- E. Primitive Chordates.- F. Vertebrates.- III. Evolution of Sialic Acids.- IV. References.- 3 The Distribution of Sialic Acids Within the Eukaryotic Cell.- I. Introduction.- II. Extracellular Sialic Acids.- III. Distribution within the Cell.- A. The Plasma Membrane.- B. Endoplasmic Reticulum.- C. Mitochondria.- D. Nuclei.- E. Other Fractions.- IV. Conclusions.- V. References.- 4 Anabolic Reactions Involving Sialic Acids.- I. Introduction: Perspective and Directions.- II. Biosynthesis of the Sialic Acids.- A. Glucose to Sialic Acid.- B. Activation.- C. Regulatory Problems.- D. Other Derivatizations.- III. Biosynthesis of Polymers, Glycoproteins, Mucins, and Glycolipids Containing Sialic Acid.- A. Colominic Acid Synthesis.- B. CMP-Sialic Acid: Lactose (?-Galactosyl) Sialyltransferase.- C. CMP-Sialic Acid: Glycoprotein (?-Galactosyl) Sialyltransferases.- D. CMP-Sialic Acid: Mucin (?-N-Acetylgalactosaminyl) Sialyltransferase.- E. CMP-Sialic Acid: Ganglioside (Glycolipid) Sialyltransferases.- IV. Thoughts on Physiological Function of Sialic Acids.- V. References.- 5 Catabolism of Sialyl Compounds in Nature.- I. Introduction.- II. Pathways of Degradation.- A. Degradation of Gangliosides.- B. Degradation of Glycoproteins.- III. Cellular Mechanism of Degradation.- A. Lysosomes.- B. Uptake and Disposition of Substrates.- IV. Functional Implications.- V. Concluding Remarks.- VI. References.- 6 Disorders of Ganglioside Catabolism.- I. Introduction-The Catabolism of Gangliosides.- II. Tay-Sachs Disease (Type I GM2-Gangliosidosis).- A. Clinical Aspects.- B. Pathology.- C. Chemistry of the Storage Material.- D. Nature of the Metabolic Defect.- E. Enzymology of Type I GM2-Gangliosidosis.- F. Prenatal Diagnosis and Treatment.- III. Type II GM2-Gangliosidosis.- A. Clinical and Pathological Aspects.- B. Chemistry of the Storage Material.- C. Metabolic Defect-Diagnosis and Treatment.- IV. Other Variant Forms.- A. Type III GM2-Gangliosidosis.- B. Hexosaminidase-A-Deficient Adults.- V. Generalized Gangliosidosis (GM1-Gangliosidosis).- A. Clinical Aspects.- B. Pathology.- C. Chemistry of the Stored Material.- D. Metabolic Defect.- VI. Potentially Related Disorders.- A. Hematoside (GM3)-Gangliosidosis.- B. Animal Model Gangliosidoses.- C. In Vitro Model Studies.- VII. References.- 7 The Biological Role of Sialic Acid at the Surface of the Cell.- I. Introduction.- II. Occurrence, Forms, and Amounts of Sialic Acid Residues at the Surface of the Cell.- III. The Masking of Cell-Surface Antigens by Sialic Acid.- IV. Sialic Acid as a Receptor at Cell Surfaces.- A. Receptor for Lectins.- B. Receptor for Viruses.- C. Receptor for Mycoplasma.- D. Receptor for Hormones.- E. Receptor for Antibodies.- F. Receptor for Circulating Glycoproteins.- G. Receptor for Tetanus Toxin.- V. Sialic Acid in Normal and Malignant or Transformed Cells.- VI. Role of Sialic Acid in Cell-to-Cell Interaction.- A. Cellular Adhesion.- B. Intercellular Aggregation.- C. Agglutination.- VII. Physiological Role of Sialic Acid Residues.- A. Transport of Ions, Amino Acids, and Proteins.- B. Phagocytosis.- C. Anaphylactic Shock, Hypercapnia, and Brain Excitability.- D. Lymphocyte Stimulation.- E. Sperm Capacitation.- VIII. Conclusion.- IX. References.- 8 The Altered Metabolism of Sialic-Acid-Containing Compounds in Tumorigenic-Virus-Transformed Cells.- I. Introduction.- II. Experimental Procedures.- A. Cells and Cell Culture.- B. Isolation, Identification, and Quantification of Gangliosides.- C. Assay of Enzymes Involved in Glycolipid Metabolism.- III. Ganglioside Metabolism in Cultured Mouse Cell Lines.- A. Distribution of Gangliosides in Normal and Virally Transformed Cells.- B. Enzymatic Studies.- C. Effect of Growth and Culture Conditions on Ganglioside Metabolism.- D. Sialic-Acid-Containing Glycolipids in Transformed Cells Obtained from Other Species.- VI. Sialic Acid and Glycoproteins in Transformed Cells.- A. Sialic Acid and Sialyltransferase Activity in Transformed Cells.- B. Membrane Glycoproteins.- C. Glycopeptides of Transformed Cells.- D. Role of Sialic Acid and Sialyltransferase.- E. Comments.- V. Relationship between Viral Transformation and Altered Ganglioside Metabolism.- A. Productive Infection of Mouse Cells.- B. Ganglioside Metabolism in Flat Revertant Cell Lines.- C. Specificity of the Altered Ganglioside Metabolism.- D. Generality of the Phenomenon.- E. Transformation of Mouse Cells by RNA Tumor Viruses and Other Agents.- VI. Discussion.- A. Molecular Basis of Altered Ganglioside Metabolism.- B. Significance.- VII. Concluding Remarks.- VIII. References.- 9 Circulating Sialyl Compounds.- I. Introduction.- II. Normal Plasma Constituents.- A. Circulating Sialoenzymes.- B. Serum Sialoglobulins.- C. Sialoglycoprotein Hormones.- III. Circulating Sialoglycoproteins in Abnormal Physiological States.- A. Diabetes.- B. Inflammatory Reactions.- C. Infectious Psychoses.- D. The Effect of Steroid Hormones.- E. Liver Disease.- F. Virus Inhibition of Hemagglutination.- G. Cancer.- H. Diet.- IV. Role of Sialic Acid in Circulating Sialoglycocompounds.- V. References.- 10 Sialidases.- I. Background and Nomenclature.- II. Bacterial Sialidases.- A. Occurrence of Microbial Sialidases.- B. Organismic Characterization and Induction of Bacterial Sialidases.- C. Purification of Bacterial Sialidases.- D. Size and Properties of Bacterial Sialidases.- E. Mode of Action of Bacterial Sialidases.- F. Biological Roles for Bacterial Sialidases.- III. Viral Sialidases.- A. Morphology and Genetics of Viral Sialidases.- B. Purification of Viral Sialidases.- C. Size of Viral Sialidases.- D. Properties of Viral Sialidases.- E. Possible Biological Roles for Viral Sialidases.- IV. Experimental Use of Microbial Sialidases.- V. Mammalian Sialidases.- A. Organ Distribution of Mammalian Sialidases.- B. Subcellular Distribution of Mammalian Sialidases.- C. Purification of Mammalian Sialidases.- D. Assay of Mammalian Sialidases.- E. Physical Properties of Mammalian Sialidases.- F. Developmental Studies of Mammalian Sialidases.- G. Possible Biological Roles of Mammalian Sialidases.- H. Sialidase Activity in Cells in Tissue Culture.- VI. References.

Botulinum Neurotoxin A Activity Is Dependent upon the Presence of Specific Gangliosides in Neuroblastoma Cells Expressing Synaptotagmin I

Botulinum neurotoxin A (BoNT/A) is the deadliest of all known biological substances. Although its toxicity makes BoNT/A a biological warfare threat, its biologic activity makes it an increasingly useful therapeutic agent for the treatment of muscular disorders. However, almost 200 years after its discovery, the neuronal cell components required for the activity of this deadly toxin have not been unequivocally identified. In this work, neuroblastoma cells expressing synaptotagmin I, a protein shown to be bound by BoNT/A, were used to determine whether specific gangliosides were necessary for BoNT/A activity as measured by synaptosomal-associated protein of 25 kDa (SNAP-25) cleavage. Ganglioside GT1b was found to support BoNT/A activity significantly more effectively than GD1a, which was far more effective than GM1 when added to ganglioside-deficient murine cholinergic Neuro 2a or to human adrenergic SK-N-SH neuroblastoma cells. Whereas both cell lines expressed synaptotagmin I, SNAP-25 cleavage was not observed in the absence of complex gangliosides. These results indicate that 1) gangliosides are required for BoNT/A activity, 2) synaptotagmin I in the absence of gangliosides does not support BoNT/A activity, and 3) Neuro 2a cells are an efficient model system for studying the biological activity of BoNT/A.

Lipid rafts: keys to neurodegeneration.

The increase in life expectancy seen in many countries has been accompanied by an increase in the number of people living with dementia and a growing need for health care. The large number of affected individuals emphasizes the need to identify causes for the phenotypes associated with diseases such as Alzheimers, Parkinsons, amyotrophic lateral sclerosis, Huntingtons, and those caused by prions. This review addresses the hypothesis that changes in lipid rafts induced by alterations in their ganglioside and/or cholesterol content or the interaction of mutant proteins with them provide the keys to understanding the onset of neurodegeneration that can lead to dementia. The biological function(s) of raft-associated gangliosides and cholesterol are discussed prior to reviewing what is known about their roles in lipid rafts in the aforementioned diseases. It concludes with some questions that need to be addressed in order to provide investigators with the basis for identifying small molecule agonists or antagonists to test as potential therapeutics.

Gangliosides: glycosphingolipids essential for normal neural development and function

Lipid rafts, sites of signal transduction, are enriched in glycosphingolipids (GSLs). Gangliosides, a class of GSLs found in greatest concentration in the grey matter of the brain, can affect neuronal function by modulating cell signaling. This review summarizes changes in ganglioside expression during brain development, the specific effects they induce, and makes observations about their possible role(s) in dementing diseases. Given that the average lifespan of individuals in many countries has increased, and that aging is accompanied by an increasing probability of dementia, understanding how changes in the GSL composition of lipid rafts may contribute to the cell biological basis of a specific dementing phenotype is an important area of study.

Oligosaccharide-derivatized dendrimers: defined multivalent inhibitors of the adherence of the cholera toxin B subunit and the heat labile enterotoxin of E. coli to GM1.

Poly(propylene imine) dendrimers having four or eight primary amino groups and a StarburstTM (PAMAM) dendrimer having eight primary amino groups were used as core molecules, to which phenylisothiocyanate derivatized (PITC) galβ1-3galNAcβ1-4[sialic acidβ2-3]-galβ1-4glc (oligo-GM1) residues were covalently attached to yield multivalent oligosaccharides. The synthesis of the oligo-GM1-PITC derivatized dendrimers was monitored using high performance thin layer chromatography, infrared spectroscopy, sialic acid content, and mass spectroscopy. The ability of multivalent oligo-GM1-PITC dendrimers to inhibit the binding of 125I-labeled cholera toxin B subunit and the heat labile enterotoxin of E. coli to GM1-coated microtiter wells was determined. IC50s obtained for the oligo-GM1-PITC dendrimers, GM1, and the oligosaccharide moiety of GM1 indicated that the derivatized dendrimers inhibited binding of the choleragenoid and the heat labile enterotoxin to GM1-coated wells at a molar concentration five- to 15-fold lower than native GM1 and more than 1,000-fold lower than that of the free oligosaccharide.

Novel Polysulfated Galactose-Derivatized Dendrimers as Binding Antagonists of Human Immunodeficiency Virus Type 1 Infection

ABSTRACT Evidence indicates that galactosyl ceramide (GalCer) and its 3′-sulfated derivative, sulfatide (SGalCer), may act as alternate coreceptors for human immunodeficiency virus type 1 (HIV-1) in CD4− cells. Glycosphingolipids (GSLs) may also be necessary for fusion of HIV-1 and host cell membranes. Using an enzyme-linked immunosorbent assay to determine which GSL was the best ligand for both recombinant and virus-associated gp120, we found that SGalCer was the best ligand for each rgp120 and HIV-1 isolate tested. Therefore, novel multivalent glycodendrimers, which mimic the carbohydrate clustering reportedly found in lipid rafts, were synthesized based on the carbohydrate moiety of SGalCer. Here we describe the synthesis of a polysulfated galactose functionalized, fifth generation DAB dendrimer (PS Gal 64mer), containing on average two sulfate groups per galactose residue. Its ability to inhibit HIV-1 infection of cultured indicator cells was compared to that of dextran sulfate (DxS), a known, potent, binding inhibitor of HIV-1. The results indicate that the PS Gal 64mer inhibited infection by the HIV-1 isolates tested as well as DxS.

Dendrimers and antivirals: a review.

In response to the need for antiviral agents, dendrimers, hyper-branched, well-defined, and chemically versatile molecules, have been found to have a number of potential uses. How they are used is based on knowledge of 1) how a virus interacts with its target cells, 2) how it replicates, and 3) which viral components are recognized by the immune response of the host. Many viral-host cell interactions are initiated by viral proteins binding to specific cell surface carbohydrates. Dendrimers offer an efficient means of presenting multiple ligands, or sites of contact, on a single molecule. Derivatized with carbohydrate residues, the multivalent ligands have been shown to inhibit viral binding. Dendrimers derivatized with peptides or anionic groups have also been found to inhibit infection. The availability of a number of different types of dendrimers permits synthesis of potential inhibitors of viral binding to be tailored to meet the dimensions needed for optimum adherence by the virus. Future directions should see increased studies of the use of dendrimers as carriers of 1) multiple indicators on a viral probe to increase diagnostic sensitivity, 2) multiple peptides for use as immunogens or as inhibitors of viral binding, and 3) inhibitors of viral enzymes. While the field of dendrimer chemistry is relatively young, promising results indicate that dendrimers may provide the scaffolding needed for development of effective antivirals.

Density-Dependent Changes in Gangliosides and Sialidase Activity of Murine Neuroblastoma Cells

Abstract: Density‐dependent changes in ganglioside composition, Vibrio cholerae neuraminidase (VCN)‐susceptible sialyl residues, and membrane‐ associated sialidase activity were determined for the cholinergic murine neuroblastoma cell line S20Y. A decrease in total ganglioside sialic acid and VCN‐releasable sialic acid was observed with increasing cell density. GM3 was the major ganglioside component of preconfluent S20Y cells, whereas GDIA was predominant in postconfluent cells. Sialidase activity increased in confluent and postconfluent cells and may account for the reduction in total ganglioside sialic acid observed with increasing cell density. In contrast, while adrenergic N115 cells showed a decrease in VCN‐susceptible sialic acid residues with increasing cell density, there was no significant change in ganglioside composition or ganglioside sialic acid levels.

Glycosphingolipids—Sweets for botulinum neurotoxin

A number of viruses, bacteria, and bacterial toxins can only act on cells that express the appropriate glycosphingolipids (GSLs) on the outer surface of their plasma membranes. An example of this dependency is provided by botulinum neurotoxin (BoNT) which is synthesized by Clostridium botulinum and inhibits neurotransmission at the neuromuscular junction by catalyzing hydrolysis of a SNARE protein, thereby inducing a flaccid paralysis. Haemagglutinin components of progenitor forms of BoNT mediate its adherence to glycosphingolipids (GSLs) on intestinal epithelial cells while the cellular activity of most isolated serotypes requires the presence of certain gangliosides, especially those of the Gg1b family. This review discusses available information about the identity and the roles of GSLs in the activity of BoNT. Observations that serotypes A-F of BoNT require gangliosides for optimum activity (serotype G apparently does not), permits the hypothesis that it should be possible to develop an antagonist of this interaction thereby inhibiting/reducing its effect. Published in 2004.

Correlation of cleavage of SNAP-25 with muscle function in a rat model of Botulinum neurotoxin type A induced paralysis

Injection of botulinum neurotoxin serotype A (BoNT/A) into muscle results in cleavage of the synaptosomal associated protein of 25 kDa (SNAP-25) and relatively long-term paralysis. However, nerve-terminal sprouting, which appears to require intact SNAP-25, has been reported to occur much earlier. The difference between the long-term paralysis induced by injection of BoNT/A and the short time needed for sprouting led us to investigate the relationship between BoNT/A catalyzed cleavage of SNAP-25 and muscle function. The effect of BoNT/A on SNAP-25 present in nerve endings innervating gastrocnemius muscles of rats was monitored over time. Cleaved SNAP-25 was found in nerve terminals innervating the muscles within 24h of inoculation with BoNT/A and was present more than 2 months later. Comparison of the ratios of cleaved to intact SNAP-25 from the onset of BoNT/A-induced paralysis until function was regained indicated that paralysis was probable when the ratio of cleaved to intact SNAP-25 was greater than 0.35.