Structures of the asparagine-linked sugar chains of laminin

This investigation describes the isolation and characterization of oligosaccharides of the basement membrane glycoprotein, laminin. Pronase-released glycopeptides of isolated laminin, from a mouse Engelbreth-Holm-Swarm tumor, were fractionated using a combination of gel permeation chromatography and...

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Vydáno v:Biochimica et biophysica acta Ročník 883; číslo 1; s. 112
Hlavní autoři: Arumugham, R G, Hsieh, T C, Tanzer, M L, Laine, R A
Médium: Journal Article
Jazyk:angličtina
Vydáno: Netherlands 06.08.1986
Témata:
Animals
Asparagine
Basement Membrane - analysis
Carbohydrate Conformation
Chromatography, Affinity
Chromatography, Gel
Galactose - analysis
Glucosamine - analysis
Glycopeptides - analysis
Laminin - analysis
Mice
Molecular Weight
Neoplasms, Experimental - analysis
Oligosaccharides - analysis
ISSN:0006-3002
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Shrnutí:This investigation describes the isolation and characterization of oligosaccharides of the basement membrane glycoprotein, laminin. Pronase-released glycopeptides of isolated laminin, from a mouse Engelbreth-Holm-Swarm tumor, were fractionated using a combination of gel permeation chromatography and Con A-Sepharose affinity chromatography. The glycopeptides were analyzed for sugar linkage patterns by methylation analysis. Glycopeptides and hydrazine-released oligosaccharides were further analyzed using endo-beta-galactosidase, endo-beta-N-acetylglucosaminidase H and specific exoglycosidases in conjunction with calibrated gel permeation chromatography. Based on these experiments, murine tumor laminin was shown to contain asparagine-linked oligosaccharides with the following structures: bi-, tri- and tetraantennary complex-type oligosaccharides; polylactosaminyl side chains containing Gal(beta 1---4)GlcNAc(beta 1---3) repeating units attached to the trimannose core portion of the bi-, tri- and tetraantennary complex-type oligosaccharides; unusual complex-type oligosaccharides terminated at the nonreducing end with sialic acid, alpha-galactose, beta-galactose and beta-N-acetylglucosamine; alpha-galactosyl residues linked to N-acetyllactosamine sequences; high-mannose-type oligosaccharides. These results, in conjunction with analytical data, indicate that most of the carbohydrate of this laminin is N-linked to asparagine and that there are about 43 such N-linked oligosaccharides per laminin molecule.
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ISSN:0006-3002
DOI:10.1016/0304-4165(86)90142-X