Document Detail

Production of procollagenase by cultured human keratinocytes.
MedLine Citation:
PMID:  2433269     Owner:  NLM     Status:  MEDLINE    
Using a collagen film assay utilizing 14C-labeled type I collagen, we demonstrated that cultured human keratinocytes produced a procollagenase after treatment with the tumor-promoting phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA). Production of collagenase paralleled alterations in cellular morphology induced by TPA. When procollagenase was immunoprecipitated with antibody to human fibroblast collagenase and analyzed on sodium dodecyl sulfate-polyacrylamide gels, the zymogen was revealed as a 56- and 51-kDa doublet. The keratinocyte-derived collagenase was a neutral metalloprotease, required activation with trypsin for detection in the collagenase assay and produced the characteristic three-quarter and one-quarter length collagen cleavage products when incubated with type I collagen at 25 degrees C. The enzyme was inhibited by serum and cysteine and was largely unaffected by serine, thiol, and carboxyl protease inhibitors. Cycloheximide inhibited the TPA-induced production of collagenase, suggesting that the procollagenase was not stored preformed in the keratinocytes. Keratinocytes treated with a tumor-promoting analogue of TPA also produced collagenase, but cells treated with cytochalasin B, interleukin-1, or two non-tumor promoting phorbol esters did not. Keratinocyte-derived collagenase may play a role in wound healing and morphogenesis.
M J Petersen; D T Woodley; G P Stricklin; E J O'Keefe
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  262     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  1987 Jan 
Date Detail:
Created Date:  1987-02-25     Completed Date:  1987-02-25     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  835-40     Citation Subset:  IM    
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MeSH Terms
Cells, Cultured
Enzyme Precursors / biosynthesis*,  metabolism
Fluorescent Antibody Technique
Infant, Newborn
Keratins / analysis*
Microbial Collagenase / biosynthesis*,  metabolism
Skin / enzymology*,  growth & development
Tetradecanoylphorbol Acetate / pharmacology
Grant Support
Reg. No./Substance:
0/Enzyme Precursors; 16561-29-8/Tetradecanoylphorbol Acetate; 68238-35-7/Keratins; EC 3.4.24.-/Collagenases; EC 3.4.24.-/procollagenase; EC Collagenase

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