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dc.contributor.authorTurnbull, Jeremy E
dc.contributor.authorGallagher, John T
dc.date.accessioned2010-11-03T15:44:56Z
dc.date.available2010-11-03T15:44:56Z
dc.date.issued1988-04-15
dc.identifier.citationOligosaccharide mapping of heparan sulphate by polyacrylamide-gradient-gel electrophoresis and electrotransfer to nylon membrane. 1988, 251 (2):597-608 Biochem. J.en
dc.identifier.issn0264-6021
dc.identifier.pmid2969727
dc.identifier.urihttp://hdl.handle.net/10541/114628
dc.description.abstractA new method that we have called 'oligosaccharide mapping' is described for the analysis of radiolabelled heparan sulphate and other glycosaminoglycans. The method involves specific enzymic or chemical scission of polysaccharide chains followed by high-resolution separation of the degradation products by polyacrylamide-gradient-gel electrophoresis. The separated oligosaccharides are immobilized on charged nylon membranes by electrotransfer and detected by fluorography. A complex pattern of discrete bands is observed covering an oligosaccharide size range from degree of polymerization (d.p.) 2 (disaccharide) to approximately d.p. 40. Separation is due principally to differences in Mr, though the method also seems to detect variations in conformation of oligosaccharide isomers. Resolution of oligosaccharides is superior to that obtained with isocratic polyacrylamide-gel-electrophoresis systems or gel chromatography, and reveals structural details that are not accessible by other methods. For example, in this paper we demonstrate a distinctive repeating doublet pattern of iduronate-rich oligosaccharides in heparitinase digests of mouse fibroblast heparan sulphate. This pattern may be a general feature of mammalian heparan sulphates. Oligosaccharide mapping should be a valuable method for the analysis of fine structure and sequence of heparan sulphate and other complex polysaccharides, and for making rapid assessments of the molecular distinctions between heparan sulphates from different sources.
dc.language.isoenen
dc.subject.meshChromatography, Gel
dc.subject.meshElectrophoresis, Polyacrylamide Gel
dc.subject.meshGlycosaminoglycans
dc.subject.meshHeparitin Sulfate
dc.subject.meshHumans
dc.subject.meshKinetics
dc.subject.meshMembranes
dc.subject.meshMethods
dc.subject.meshNitrous Acid
dc.subject.meshNylons
dc.subject.meshOligosaccharides
dc.subject.meshPolysaccharide-Lyases
dc.titleOligosaccharide mapping of heparan sulphate by polyacrylamide-gradient-gel electrophoresis and electrotransfer to nylon membrane.en
dc.typeArticleen
dc.contributor.departmentDepartment of Clinical Research, University of Manchester, Christie Hospital and Holt Radium Institute, U.K.en
dc.identifier.journalThe Biochemical Journalen
html.description.abstractA new method that we have called 'oligosaccharide mapping' is described for the analysis of radiolabelled heparan sulphate and other glycosaminoglycans. The method involves specific enzymic or chemical scission of polysaccharide chains followed by high-resolution separation of the degradation products by polyacrylamide-gradient-gel electrophoresis. The separated oligosaccharides are immobilized on charged nylon membranes by electrotransfer and detected by fluorography. A complex pattern of discrete bands is observed covering an oligosaccharide size range from degree of polymerization (d.p.) 2 (disaccharide) to approximately d.p. 40. Separation is due principally to differences in Mr, though the method also seems to detect variations in conformation of oligosaccharide isomers. Resolution of oligosaccharides is superior to that obtained with isocratic polyacrylamide-gel-electrophoresis systems or gel chromatography, and reveals structural details that are not accessible by other methods. For example, in this paper we demonstrate a distinctive repeating doublet pattern of iduronate-rich oligosaccharides in heparitinase digests of mouse fibroblast heparan sulphate. This pattern may be a general feature of mammalian heparan sulphates. Oligosaccharide mapping should be a valuable method for the analysis of fine structure and sequence of heparan sulphate and other complex polysaccharides, and for making rapid assessments of the molecular distinctions between heparan sulphates from different sources.


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