Type III procollagen and collagen in skin.
A form of collagen, containing three alpha chains of type III, can be extracted from foetal calf, calf and rat skin under physiological conditions. This native collagen was purified by DEAE-cellulose chromatography and then was analysed by polyacrylamide gel electrophoresis which showed it consisted of several high molecular weight components, the size of gamma components and larger species. Prior reduction in dithiothreitol dissociated these large polymers into two components: the minor one migrated between the alpha1 (I) and alpha2 chains while the predominant one migrated between the alpha and beta chains. These two monomers were isolated by CM-cellulose chromatography. The minor one, which eluted between the alpha1 and alpha2 chains, had a molecular weight of approx. 95 000; its amino acid composition was similar to that of alpha1(III). The major one eluted in the alpha1 region and had a molecular weight of approx. 120 000; its amino acid composition, while similar to that of the alpha1(III) chain, differed in detail, and it is presumed to be a pro-alpha1(III) chain. Following pepsin digestion, the native collagen remained as a disulfide-bonded trimer which dissociated into only one component, a1(III), when denatured in dithiothreitol. These results suggest that the original, extracted protein consisted primarily ofa precursor form of type III collagen. This procollagen did not polymerize when heated at 37 degrees C and did not form the usual segment long spacing aggregates under suitable conditions. It was not modified by incubation with a purified procollagen peptidase preparation. This appears to be the first example of the isolation of type III (pro)collagen by extractive methods, without resorting to tissue digestion by proteolytic enzymes.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D002848:Chromatography, DEAE-Cellulose; D002852:Chromatography, Ion Exchange; D003094:Collagen / Q000032:analysis; D004220:Disulfides; D004229:Dithiothreitol; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005333:Fetus; D046911:Macromolecular Substances; D008970:Molecular Weight; D010434:Pepsin A; D011487:Protein Conformation; D011498:Protein Precursors / Q000032:analysis; D051381:Rats; D012867:Skin / Q000032:analysis*; D013696:Temperature; D014783:Viscosity
D000596:Amino Acids; D004220:Disulfides; D046911:Macromolecular Substances; D011498:Protein Precursors; D003094:Collagen; D010434:Pepsin A; D004229:Dithiothreitol