Characterization of collagen precursors found in rat skin and rat bone.
Two genetic types of collagenous proteins, type I and type III, were isolated by extraction and differential salt precipitation from rat skin. The yield of collagen precursors was increased by injecting animals with colchicine 30 min before sacrifice to inhibit secretion of collagen. DEAE-cellulose chromatography was used to separate collagen from collagen precursors. Although these preparations contained more type I collagen than type III collagen, there were always more type III than type I precursors. The precursor chains of type I fractions were separated on CM-cellulose chromatography after denaturation. Three precursor forms were found for each collagen alpha chain, a complete chain (proalpha chain), and a precursor chain with only an amino-terminal (pNalpha chain) and carboxy-terminal extension (pCalpha chain). Species differences were demonstrated between rat collagen precursors and other species using rat calvaria (frontal and parietal) bones extracted with either 0.5 N acetic acid or neutral salt buffers containing protease inhibitors. Native rat procollagen elutes earlier than chicken or human procollagen on DEAE-cellulose chromatography and does not separate significantly from the pC collagen form. The collagenase resistant amino terminal peptides of rat pNalpha1 and pNalpha2 were the same size (16 000) but could be separated by DEAE-cellulose chromatography.
D000818:Animals; D000831:Animals, Newborn; D001842:Bone and Bones / Q000187:drug effects / Q000378:metabolism*; D003078:Colchicine / Q000494:pharmacology; D003012:Microbial Collagenase; D011347:Procollagen / Q000096:biosynthesis* / Q000302:isolation & purification; D011392:Proline / Q000494:pharmacology; D051381:Rats; D012867:Skin / Q000187:drug effects / Q000378:metabolism*; D012886:Skull
D011347:Procollagen; D011392:Proline; D003012:Microbial Collagenase; D003078:Colchicine