Human skin collagenase: isolation of precursor and active forms from both fibroblast and organ cultures

Abstract

Human skin procollagenase has been isolated, in pure form, from the medium of fibroblasts cultured in the presence or absence of added serum. Purification was achieved using a combination of cation-exchange (phosphocellulose or carboxymethylcellulose) and gel-filtration chromatography. Two forms (60 000 and 55 000 daltons) of the procollagenase were detected by electrophoresis in sodium dodecyl sulfatepolyacrylamide gels and could be separated by chromatography on Ultrogel AcA-44. Each form was converted to active enzyme by trypsin, producing species of 50 000 and 45 000 daltons, respectively. An autoactivation process also occurred, which yielded active enzyme without a detectable change in molecular weight. Procollagenase also was found in organ cultures of human skin but only when serum was added to the medium. This suggests that a serum-inhibitable proteolytic system is present in these cultures which, like trypsin, converts procollagenase to the active enzyme forms that can be isolated from serum-free organ culture medium. The collagenase species obtained from either fibroblast or organ culture medium were chromatographically and electrophoretically identical.

Influence of LPS, TNF, TGF-�1 and IL-4 on the expression of MMPs, TIMPs and selected cytokines in rat synovial membranes incubated in vitro

“…MMP1a is able to degrade type I, II and III fibrillar collagen (99,100). It is produced by fibroblasts as procollagenase (101) and is activated by MMP3 (stromelysin) (102). Its expression in human synovial fibroblasts has been shown to be stimulated by TNF (103).…”

Binding of plasminogen to Escherichia coli adhesion proteins

“…Binding of plasminogen to bacterial fimbriae may thus generate localized plasmin activity on the bacterial surface. Since plasmin effectively digests different extracellular matrix proteins [28,29] and activates collagenase [30], bacterium-bound plasmin may provide an effective means for bacteria to invade host tissues. On the other hand, the present finding that plasmin treatment of type 1 fimbriae did not influence their ability to agglutinate yeast ceils suggests that fimbriae themselves are not digested by plasmin.…”

Purification and Characterization of a Collagenolytic Protease from the Filefish, Novoden modestrus

“…Metallo-collagenases, first discovered in tadpole tissue explants (Gross and Lapiere, 1962), are zinc-containing enzymes that also generally require calcium for their optimum activity and stability, and cleave the collagen helix at a specific locus under physiological conditions (Gawston and Murphy, 1981;Harris and Vater, 1982;Sellers and Murphy, 1981;Stricklin et al, 1977). These enzymes have been widely studied from various mammalian tissues (Harris and Vater, 1982;Sellers and Murphy, 1981) as well as from bacteria, such as Bacillus cereus (Makinen and Makinen, 1987), Clostridium histolyticum (Bond and Van Wart, 1984a, b;Peterkofsky, 1982), Achromobacter (Nguyen et al, 1988), Vibrio alginolyticus (Takeuchi et al, 1992) and Clostridium perfringens (Matsuhita et al, 1994), and snake venom (Bjarnason and Fox, 1994).…”