Abstract
Secreted as well as membrane-associated eukaryotic proteins are most commonly glycosylated. Saccharides are attached to proteins mainly through N- and O-glycosydic bonds or as part of the glycosylphosphatidyinositolmembrane anchor. In contrast to N-glycosylation, which involves the co-translational transfer in the endoplasmic reticulum (ER) of the glycan portion of Glc3Man9GlcNAc2-PP-dolichol to suitable Asn residues on nascent polypeptides, O-glycosylation begins with the addition of a single monosaccharide. Contrary to N-glycosylation, which involves an asparagine residue in the sequon Asn-Xaa-Thr/Ser (Xaa can be any amino acid except Pro, and it is rarely Cys), no particular sequence motif has been described for O-glycosylation. This may reflect the fact that: (1) the specificity of the UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase is presently unknown; and (2) seems to be modulated by sequence context, secondary structure, and surface accessibility (1). An internet server, accessible at http://www.cbs.dtu.dk/netOglyc/cbsnetOglyc.html, produces neural network predictions of mucin-type GalNAc O-glycosylation sites in mammalian proteins based on 299 known and verified mucin-type O-glycosylation sites. The sequence context of glycosylated threonines was found to differ from that of serine, and the sites were found to cluster. Nonclustered sites had a sequence context different from that of clustered sites, and charged residues were disfavored at position −1 and +3.
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References
Hansen J. E., Lund O., Tolstrup N., Gooley A. A., Williams K. L., and S. Brunak S. (1998) NetOglyc: Prediction of mucin type O-glycosylation sites based on sequence context and surface accessibility. Glycoconjugate J. 15, 115–130.
van den Steen P., Rudd P. M., Dwek R. A., and Opdenakker G. (1998) Concepts and principles of O-linked glycosylation. Crit. Rev. Biochem. Mol. Biol. 33, 151–208.
Gemmill T. R. and Trimble R. B. (1999) Overview of N-and O-linked oligosaccharide structures in various yeast species. Biochim. Biophys. Acta 1426, 227–237.
Strahl-Bolsinger S., Gentzsch M., and Tanner W. (1999) Protein O-mannosylation. Biochim. Biophys. Acta 1426, 297–307.
Hounsell E. F., Davies M. J., and Renouf D. V. (1996) O-linked protein glycosylation structure and function. Glycoconjugate J. 13, 19–26.
Fountoulakis M. and Lahm H-W. (1998) Hydrolysis and amino acid composition of proteins. J. Chromatogr. A 826, 109–134.
Whittal R. M., Palcic M. M., Hindsgaul O., and Li L. (1995) Direct analysis of enzymatic reactions of oligosaccharides in human serum using matrix-assisted laser desorption ionization mass spectrometry. Anal. Chem. 67, 3509–3514.
Eirín M. T., Calvete J. J., and González-odríguez J. (1986) New isolation procedure and further biochemical characterization of glycoproteins IIb and IIIa from human platelet plasma membrane. Biochem. J. 240, 147–153.
Calvete J. J. and González-Rodríguez J. (1986) Isolation and biochemical characterization of the α-and β-subunits of glycoprotein IIb of human platelet plasma membrane. Biochem. J. 240, 155–161.
Montreuil J., Bouquelet S., Debray H., Lemoine J., Michalski J-C., Spik G., and Strecker G. (1994) Glycoproteins, in Carbohydrate Analysis. A Practical Approach (Chaplin M. F. and Kennedy J. F., eds.), IRL Press, Oxford, pp. 181–293.
Dell A., Khoo K-H., Panico M., McDowell R. A., Etienne A. T., Reason A. J., and Morris H. R. (1993) FAB-MS and ES-MS of glycoproteins, in Glycobiology. A Practical Approach (Fukuda M. and Kobata A., eds.), IRL Press, Oxford, pp. 187–222.
Alving K., Paulsen H., and Peter-Katalinic J. (1999) Characterization of O-glycosylation sites in MUC2 glycopeptides by nanoelectrospray QTOF mass spectrometry. J. Mass Spectrom. 34, 395–407.
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Calvete, J.J., Sanz, L. (2002). Analysis of O-Glycosylation. In: Kannicht, C. (eds) Posttranslational Modifications of Proteins. Methods in Molecular Biology™, vol 194. Humana Press. https://doi.org/10.1385/1-59259-181-7:089
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DOI: https://doi.org/10.1385/1-59259-181-7:089
Publisher Name: Humana Press
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