title: Cell-layer-associated proteolytic cleavage of the telopeptides of type I collagen in fibroblast culture.
abstract: In human skin fibroblast cultures a fraction of the procollagen that was processed to collagen and remained in the cell layer was further proteolytically modified by removal of both N- and C-terminal telopeptides. The proteolytic activity was associated with the cell layer, since secreted collagens were found always to contain intact telopeptides. The inclusion of neutral polymers, which caused the accumulation of the collagen in the cell layer [Bateman, Cole, Pillow & Ramshaw (1986) J. Biol. Chem. 261, 4198-4203], made the telopeptide cleavage more apparent in those cells which expressed the proteolytic activity. The extent of this cleavage was variable from cell culture to cell culture and between experiments with the same fibroblast line. The proteolytic activity was pH-dependent; cleavage was greatest at a culture-medium pH of 7.5 and 8.0 and was completely inhibited at a culture-medium pH of 7.0 and 6.5. The activity was significantly inhibited by soybean trypsin inhibitor, an elastase-specific inhibitor (N-acetylalanylalanylprolylvalylchloromethane) and the thrombin inhibitor hirudin. This cell-associated proteolytic activity may play a role in collagen degradation by removing the telopeptides, which are the primary sites of collagen cross-linking, thus destabilizing the collagen matrix sufficiently to render it susceptible to further proteolytic breakdown.
authors.affiliation: Department of Paediatrics, University of Melbourne, Royal Children's Hospital, Parkville, Australia.
authors.forename: J F
authors.initials: JF
authors.lastname: Bateman
authors.forename: J J
authors.initials: JJ
authors.lastname: Pillow
authors.forename: T
authors.initials: T
authors.lastname: Mascara
authors.forename: S
authors.initials: S
authors.lastname: Medvedec
authors.forename: J A
authors.initials: JA
authors.lastname: Ramshaw
authors.forename: W G
authors.initials: WG
authors.lastname: Cole
mesh_terms: D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005109:Extracellular Matrix / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D010447:Peptide Hydrolases / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism*; D011205:Povidone / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology
chemical_list: D010455:Peptides; D011480:Protease Inhibitors; D003094:Collagen; D010447:Peptide Hydrolases; D011205:Povidone