Identification of a large interrupted helical domain of disulfide-bonded cartilage collagen.

In order to characterize a larger form of disulfide-bonded cartilage collagen, explants of 17-day embryonic chick sterna were cultured in the presence of [3H] proline. Radioactive collagen chains and fragments that were synthesized and secreted into the culture medium were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography. After limited pepsin digestion of the medium, two discrete disulfide-bonded collagen fragments were detected with Mr = 210,000 and 153,000. These fragments contained 28 and 17.5%, respectively, of the radioactivity in the alpha 1(II)-chains. The smaller fragment (called M) produced three components upon reduction (Mr = 104,000, 51,000, and 31,000) and seemed to represent the previously reported collagens, HMW and M1. The larger fragment (called N) has not been previously described and gave rise to three components upon reduction (Mr = 140,000, 69,000, and 49,000). Prolonged pepsin treatment resulted in the gradual decrease of N with a corresponding increase of M, suggesting the conversion of N to M. CNBr peptide mapping demonstrated that all M-derived peptides were present in N and that N contained extra peptides that account for its larger size. These observations suggest that N represents a larger more intact form of cartilage-derived disulfide-bonded collagen.