| Abstract: | In cultures of dermal fibroblasts, procollagen and the intermediates pC- and pN-collagen accumulated in the culture medium with little further processing to collagen. When polyethylene glycol (PEG) or other neutral polymers were added to fibroblast culture medium, no collagen or procollagen was found in the medium, but all the collagen was associated with the cell layer. The type I procollagen was fully processed to collagen with an initial transient accumulation of pN-collagen, and the processed collagen formed covalently cross-linked dimers. The presence of pepsin-sensitive COOH-terminal telopeptides and the accumulation of pN-collagen in PEG-treated cultures of dermatosparactic fibroblasts indicated that it was likely that processing occurred via the correct in vivo propeptidase activities. At the levels used in this study, PEG did not have any toxic effect during the incubation period on the fibroblasts in culture, since the amount of total protein synthesis was not altered by addition of PEG to cultures. However, the level of collagen production was reduced to about half, indicating that there was increased degradation or that the released collagen propeptides or the accumulation of processed collagen in association with the cells exerted a feedback regulation on collagen synthesis. Addition of neutral polymers to the culture medium provided a simple means of achieving complete and accurate processing of procollagen which more closely resembled the in vivo process. |
