title: Effect of tunicamycin on the biosynthesis of human fibroblast collagenase.
abstract: It is generally accepted that collagenase from human fibroblast cultures consists of two proenzymes (Mr 60,000 and 55,000) and two active forms (Mr 50,000 and 43,000). We demonstrated previously that epidermal growth factor (EGF) as well as a number of other growth factors induced the secretion of procollagenase (Mr 60,000, Mr 55,000) into the medium of human fibroblast cultures (Chua et al., 1985). In the presence of tunicamycin and EGF, the secretion of the larger form of procollagenase was suppressed preferentially with concomitant appearance of a new band, Mr 40,000. This Mr 40,000 band could be specifically immunoprecipitated by antibody raised against human collagenase. By two-dimensional peptide mapping, the Mr 40,000 material appeared to have similar composition as the Mr 60,000 band. In a time course study, the Mr 55,000 procollagenase band was the earliest protein to appear in the medium after 1 hour labeling with [35S]-methionine. The Mr 60,000, 50,000 and 43,000 bands appeared after a 2 hour labeling period. Our results indicate that human collagenases are glycosylated proteins and are synthesized via the dolichol phosphate pathway.
authors.affiliation: Department of Immunology and Microbiology, Wayne State University School of Medicine, Detroit, MI 48201.
authors.forename: C C
authors.initials: CC
authors.lastname: Chua
authors.forename: R L
authors.initials: RL
authors.lastname: Ladda
mesh_terms: D017364:Collagenases; D004591:Electrophoresis, Polyacrylamide Gel; D004792:Enzyme Precursors / Q000096:biosynthesis; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007163:Immunosorbent Techniques; D003012:Microbial Collagenase / Q000096:biosynthesis*; D008970:Molecular Weight; D014415:Tunicamycin / Q000494:pharmacology*
chemical_list: D004792:Enzyme Precursors; D014415:Tunicamycin; D004815:Epidermal Growth Factor; D017364:Collagenases; C022440:procollagenase; D003012:Microbial Collagenase