Sialidases

Leland D. Powell1, Ajit P. Varki1

1 University of California San Diego, La Jolla, California
Publication Name:  Current Protocols in Molecular Biology
Unit Number:  Unit 17.12
DOI:  10.1002/0471142727.mb1712s27
Online Posting Date:  May, 2001
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Abstract

Sialic acids are a family of nine‐carbon acidic sugars found at the nonreducing terminus of many glycoconjugates. Sialidases can remove these sugar units selectively from cell surfaces, membranes, or purified glycoconjugates. In this unit, sialidase digestion of purified glycoproteins is described as is treatment of intact cells. The physical properties of the four most useful sialidases are discussed along with their relative activities against sialic acids with different modifications and in different linkages

     
 
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Table of Contents

  • SECTION III: Release of Saccharides from Glycoconjugates
  • Basic Protocol 1: Sialidase Treatment of Purified Glycoproteins
  • Alternate Protocol 1: Sialidase Treatment of Intact Cells
  • Reagents and Solutions
  • Commentary
  • Tables
     
 
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Materials

Basic Protocol 1: Sialidase Treatment of Purified Glycoproteins

  Materials
  • Sialic acid–containing sample
  • recipeSialidase digestion buffer
  • Sialidase (Table 17.12.1)
  • Additional reagents and equipment for quantitating sialic acid (unit 17.16)

Alternate Protocol 1: Sialidase Treatment of Intact Cells

  Additional Materials
  • Cells, prepared as a single‐cell suspension
  • recipeHEPES‐buffered saline (HeBS)
  • Culture medium appropriate for cells, with serum
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Literature Cited

Literature Cited
   Ada, G.L., French, E.L., and Lind, P.E. 1961. Purification and properties of neuraminidase from Vibrio cholerae. J. Gen. Microbiol. 24:409‐421.
   Cassidy, J.T., Jourdian, G.W., and Roseman, S. 1965. The sialic acids IV: Purification and properties of sialidase from Clostridium perfringens. J. Biol. Chem. 240:3501‐3506.
   Drzeniek, R. 1973. Substrate specificity of neuraminidases. Histochem. J. 5:271‐290.
   Townsend, R.R., Hardy, M.R., Cumming, D.A., Carver, J.P., and Bendiak, B. 1989. Separation of branched sialylated oligosaccharides using HPAE‐PAD. Anal. Biochem. 182:1‐8.
   Troy, F.A. 1992. Polysialylation: From bacteria to brains. Glycobiology 2:5‐24.
   Uchida, Y., Tsukada, Y., and Sugimori, T. 1979. Enzymatic properties of neuraminidases from Arthrobacter ureafaciens. J. Biochem. 86:1573‐1585.
   Varki, A. 1992. Diversity in the sialic acids. In Glycobiology Vol. 2 (1): 25‐40.
Key References
   Corfield, A.P., Wember, M., Schauer, R., and Rott, R. 1982. The specificity of viral sialidases. Eur. J. Biochem. 124:521‐525.
  Describes biochemical properties of the Newcastle disease virus sialidase.
   Corfield, A.P., Higa, H.H., Paulson, J.C., and Schauer, R. 1983. The specificity of viral and bacterial sialidases for alpha (2‐3) and alpha (2‐6) linked sialic acids in glycoproteins. Biochim. Biophys. Acta 744:121‐126.
  Provides characterization of sialidases on α(2‐3) and α(2‐6) linked sialic acids.
   Corfield, A.P., Sander Wewer, M., Veh, R.W., Wember, M., and Schauer, R. 1986. The action of sialidases on substrates containing O‐acetyl sialic acids. Biol. Chem. Hoppe‐Seyler 367:433‐439.
  Describes the activity of different sialidases on O‐acetylated sialic acids.
   Varki, A. and Diaz, S. 1984. The release and purification of sialic acids from glycoconjugates: Methods to minimize the loss and migration of O‐acetyl groups. Anal. Biochem. 137:236‐247.
  Describes essential steps necessary to preserve O‐acetyl groups during release and characterization of sialic acids.
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