Separation of radiolabelled glycosaminoglycan oligosaccharides by polyacrylamide-gel electrophoresis.
AbstractGlycosaminoglycan oligosaccharides generated by treatment of biosynthetically radiolabelled dermatan sulphate and hyaluronic acid with chondroitin AC lyase or testicular hyaluronidase may be resolved into a series of discrete bands by polyacrylamide-gel electrophoresis. Bands were identified by fixation in glacial acetic acid containing 20% (w/v) 2,5-diphenyloxazole followed by fluorography. The bands represented glycans which differed in size by one disaccharide unit. For the larger oligosaccharides (decasaccharides and above) of similar charge: mass ratio, there was a linear relationship between electrophoretic mobility and log Mr. However, the smaller species showed anomalous migration patterns. Consideration of the structures of the fragments produced by the different enzyme treatments suggests that copolymeric and homopolymeric oligosaccharides may be separated by polyacrylamide-gel electrophoresis. There are many potential applications of this technique, foremost amongst them being studies on the molecular size heterogeneity and patterns of enzyme-mediated depolymerization of native glycosaminoglycan chains and investigations into rates of polymer chain elongation and post-polymerization modification reactions so essential to glycosaminoglycan function.
CitationSeparation of radiolabelled glycosaminoglycan oligosaccharides by polyacrylamide-gel electrophoresis. 1984, 221 (3):697-705 Biochem J
JournalThe Biochemical Journal
- Polyacrylamide-gel electrophoresis and Alcian Blue staining of sulphated glycosaminoglycan oligosaccharides.
- Authors: Cowman MK, Slahetka MF, Hittner DM, Kim J, Forino M, Gadelrab G
- Issue date: 1984 Aug 1
- Oligosaccharide mapping of heparan sulphate by polyacrylamide-gradient-gel electrophoresis and electrotransfer to nylon membrane.
- Authors: Turnbull JE, Gallagher JT
- Issue date: 1988 Apr 15
- A general method for the detection and mapping of submicrogram quantities of glycosaminoglycan oligosaccharides on polyacrylamide gels by sequential staining with azure A and ammoniacal silver.
- Authors: Lyon M, Gallagher JT
- Issue date: 1990 Feb 15
- Formation of dermatan sulfate by cultured human skin fibroblasts. Effects of sulfate concentration on proportions of dermatan/chondroitin.
- Authors: Silbert JE, Palmer ME, Humphries DE, Silbert CK
- Issue date: 1986 Oct 15
- Application of fluorophore-assisted carbohydrate electrophoresis to analysis of disaccharides and oligosaccharides derived from glycosaminoglycans.
- Authors: Oonuki Y, Yoshida Y, Uchiyama Y, Asari A
- Issue date: 2005 Aug 15