Collagenases from human and rat skin fibroblasts purified on a zinc chelating column reveal marked differences in latency as a result of serum culture conditions.
1. Fibroblasts from both human and rat skin were grown in the presence or absence of serum and the collagenase activity in the medium was partially purified on zinc-Sepharose. 2. During chromatography, using a discontinuous elution gradient, the rat collagenase elutes at different pH and ionic strength than the human collagenase. Both latent and active collagenases of both species are retarded by the affinity matrix. 3. Latency of collagenase in media obtained from fibroblast cultures appears to be influenced by the presence of a serum component in the culture medium. 4. The results demonstrate that collagenases secreted by fibroblast cultures established from the same tissue but obtained from different species are biochemically diverse and that, within one species, the amount of active enzyme depends on the presence of a serum factor.
van der Stappen
J W
JW
Department of General Surgery, St Radboud University Hospital, Nijmegen, The Netherlands.
Hendriks
T
T
de Man
B M
BM
D000818:Animals; D001769:Blood; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology*; D006801:Humans; D003012:Microbial Collagenase / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D015032:Zinc
D003470:Culture Media; D003012:Microbial Collagenase; D015032:Zinc