Bovine glomerular basement membrane. Characterization of an alpha-size collagenous polypeptide.
The collagenase domain of bovine glomerular basement membrane was isolated in soluble form after limited digestion with pepsin. Gel filtration chromatography of the domain under denaturing conditions revealed that most of the polypeptide constituents exhibit apparent molecular weights greater than the type I collagen beta-chain, while approximately 15% are similar in size to that of alpha-chain. Carboxymethyl cellulose chromatography of the alpha-size region revealed that 70% of the protein was polypeptide XIV, as previously designated (West, T. W., Fox, J. W., Jodlowski, M., Freytag, J. W., and Hudson, B. G. (1980) J. Biol. Chem. 255, 10451-10459). This polypeptide exhibits an apparent molecular weight of 102,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. An absolute molecular weight value of 86,000 was determined by sedimentation equilibrium ultracentrifugation in 6 M guanidine hydrochloride. About 15% of the mass is carbohydrate which exists in the form of glucosylgalactosylhydroxylysine. Thus, the polypeptide backbone has a molecular weight of 73,000, a value which is considerably smaller than the alpha-chains of classical collagen. The amino acid and carbohydrate composition and cyanogen bromide patterns indicate that polypeptide XIV has a structure similar to that of C-chain or alpha 1 (IV) collagen which has been identified in other tissues. In addition, the cyanogen bromide pattern of the entire collagenous domain is similar to that of polypeptide XIV, suggesting that the latter is a structural segment of many of the higher molecular weight components.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001485:Basement Membrane / Q000032:analysis*; D002241:Carbohydrates / Q000032:analysis; D002417:Cattle; D003094:Collagen / Q000032:analysis*; D003488:Cyanogen Bromide; D007678:Kidney Glomerulus / Q000032:analysis*; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis; D010455:Peptides / Q000032:analysis*
D000596:Amino Acids; D002241:Carbohydrates; D010446:Peptide Fragments; D010455:Peptides; D003094:Collagen; D003488:Cyanogen Bromide