Characterization of procollagen synthesized by matrix-free cells isolated from chick embryo tendons.
The genetic type and molecular structure of the precursor forms of collagen synthesized by matrix-free tendon cells isolated from 17-day old chick embryos were examined by chromatographic and electrophoretic techniques. The [14C]proline-labeled collagenous proteins secreted by the cells resolved on diethylaminoethylcellulose into two peaks, A and B. Both peaks contained type I collagenous proteins since on chromatography on carboxymethylcellulose, after limited pepsin proteolysis, both peaks contained alpha1 and alpha2 chains of collagen in a 2:1 ratio, and cyanogen bromide peptide maps of the 14C-labeled protein in both peaks were similar to cyanogen bromide peptide maps derived from authentic type I collagen. Enzymatic digestion with purified mammalian collagenase demonstrated that the collagen precursor in peak B contained noncollagenous peptide extensions at both the amino- and carboxy-terminal ends of the molecule, while peak A had only carboxy-terminal extension peptides. Although both the amino- and carboxy-terminal extensions incorporated radioactive cystine, only the carboxy-terminal extensions contained interchain disulfide bonds. The carboxy-terminal extensions were also shown to incorporate radioactive tryptophan. Since most of the precursor forms of collagen recovered in the incubation medium chromatographed in peak B, it is concluded that matrix-free tendon cells secrete only type I procollagen with extension peptides at both the amino- and carboxy-terminal ends of the molecule.
D000818:Animals; D002642:Chick Embryo; D006801:Humans; D066298:In Vitro Techniques; D046911:Macromolecular Substances; D003012:Microbial Collagenase / Q000302:isolation & purification; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis; D011347:Procollagen / Q000096:biosynthesis / Q000302:isolation & purification; D012867:Skin / Q000201:enzymology; D013710:Tendons / Q000378:metabolism*
D046911:Macromolecular Substances; D010446:Peptide Fragments; D011347:Procollagen; D003012:Microbial Collagenase