Glycoproteins: rapid sequencing technology for N-linked and GPI anchor glycans

Journal article

Recent advances in oligosaccharide sequencing technology have made routine glycan analysis of glycoproteins and the glycan moieties of glycosylphosphatidylinositol (GPI) anchors a real possibility for many laboratories. The strategies described here have been developed specifically to address the need for a rapid and robust method of glycan analysis which can be applied routinely to microgram levels of glycoproteins with the minimum requirement for specialised equipment or expertise. This strategy also allows the possibility of isolating individual sugars of particular interest for more detailed analysis. We discuss the N~glycosy[ation ofCD48. IgO and IgA I, the Fab and Fc fragments of [gO and also the small (S) and middle (M) glycoproteins of the hepatitiS B virus coat protein. We describe the analysis of the major O·glycans of neutrophil gelatinase B and also present a novel method for analysing the gIycans attached to the GPI anchor of a variant surface glycoprotein of Trypcl1Z0S()mlllmlcei directly from a Western blot. The underlying aim of these analytical strategies is to obtain oligosaccharide sequencing data which. in combination with oligosaccharide and protein structural data. can be visualised in a molecular model. In this way, oligo saccharides can be viewed in the context of the proteins to which they are attached. and some insight can be gained into the roles which sugars might play in the structure and function of the glycoproteins.

Authors: Rudd, P; Mattu, T; Zitzmann, N; Mehta, A; Colominas, C

• Publication status: Published
• Peer review status: Peer reviewed
• Publisher: Taylor and Francis
• Journal: Biotechnology and Genetic Engineering Reviews
• Volume: 16
• Issue: 1
• Pages: 1-21
• Publication date: 1999-04-01
• DOI: 10.1080/02648725.1999.10647969
• EISSN: 2046-5556
• ISSN: 0264-8725
• Language: English
• Keywords: biotechnology; glycoproteins; chromatography, high pressure liquid; immunoglobulin G; sequence analysis; glycosylation; glycosylphosphatidylinositols; immunoglobulin A; carbohydrate conformation; models, molecular; molecular sequence data; carbohydrate sequence; protein structure, tertiary; polysaccharides; oligosaccharides