Radioactive tracer techniques in the sequencing of glycoprotein oligosaccharides.
Complete sequencing of the oligosaccharide units of glycoproteins can be performed by conventional physical techniques if nanomole quantities of pure molecules are available. However, isolation of sufficient quantities of a glycoprotein may not be technically feasible (e.g., the analysis of biosynthetic intermediates, or rare molecules). Alternatively, partial structural analyses may answer the biological question at hand. In both instances, radioactive sugars can be used to metabolically label the oligosaccharide units of a glycoprotein, permitting substantial structural characterization. Several aspects of this approach are discussed in this overview, including selection of the labeled precursor, maximization of uptake and incorporation, determinants of the specificity of labeling, and general principles for the release and structural analysis of labeled oligosaccharides. Particular advantages include simplicity, ease of use without sophisticated instrumentation, and the fact that purification to radiometric homogeneity is sufficient. Radioactive tracer techniques cannot replace conventional approaches to sequencing oligosaccharides. However, they do provide a rapid, relatively simple approach to obtaining considerable information from limited amounts of material. For molecules such as short-lived biosynthetic intermediates, there is no substitute for these techniques. This approach has been responsible for the initial identification and characterization of many novel oligosaccharides of biological interest.
Varki
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Cancer Biology Program, UCSD Cancer Center.
D000818:Animals; D002240:Carbohydrate Sequence; D002846:Chromatography, Affinity; D006001:Glycoconjugates / Q000096:biosynthesis / Q000737:chemistry*; D006020:Glycopeptides / Q000737:chemistry; D006023:Glycoproteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D006026:Glycoside Hydrolases / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D037102:Lectins; D009005:Monosaccharides / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D011849:Radioactive Tracers
D006001:Glycoconjugates; D006020:Glycopeptides; D006023:Glycoproteins; D037102:Lectins; D009005:Monosaccharides; D011849:Radioactive Tracers; D006026:Glycoside Hydrolases