Different levels of glycosylation contribute to the heterogeneity of alpha 1(II) collagen chains derived from a transplantable rat chondrosarcoma.
Three collagen fractions, each of which contain molecules composed of alpha 1(II) chains, have been isolated from pepsin-solubilized rat chondrosarcoma collagen. One fraction could be selectively precipitated from the pepsin digest at 0.7 M NaCl. Two additional fractions were obtained on chromatography of the collagen precipitating at 1.2 M NaCl on carboxymethyl cellulose under nondenaturing conditions. When chromatographed on carboxymethyl cellulose under denaturing conditions, each fraction contained components eluting in the position expected for alpha 1(II) chains. One of the fractions precipitating at 1.2 M NaCl contained the recently described 1 alpha and 2 alpha chains in addition to material eluting as alpha 1(II) chains. Comparison of the chains eluting as alpha 1(II) chains in the various fractions with respect to amino acid composition, carbohydrate content, and cyanogen bromide-cleavage products showed that they differed only in the number of glycosylated hydroxylysyl residues. In this regard, alpha 1(II) chains obtained from collagens precipitated at 1.2 M NaCl exhibited significantly higher levels of glucosylgalactosylhydroxylysyl residues than alpha 1(II) chains precipitated at 0.7 M NaCl. These results indicate that molecules composed of alpha 1(II) chains are heterogeneous with respect to levels of hydroxylysine-linked carbohydrate moieties and that the more highly glycosylated molecules require higher salt concentrations for precipitation from acidic solutions. The data also indicate that a proportion of the more highly glycosylated alpha 1(II) chains are involved in the formation of one or more molecular species with 1 alpha and 2 alpha chains.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002460:Cell Line; D002813:Chondrosarcoma / Q000378:metabolism*; D003094:Collagen / Q000235:genetics* / Q000302:isolation & purification; D003488:Cyanogen Bromide; D006027:Glycosides / Q000378:metabolism; D046911:Macromolecular Substances; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis; D011499:Protein Processing, Post-Translational; D051381:Rats
D000596:Amino Acids; D006027:Glycosides; D046911:Macromolecular Substances; D010446:Peptide Fragments; D003094:Collagen; D003488:Cyanogen Bromide