The latent collagenase and gelatinase of human polymorphonuclear neutrophil leucocytes

Abstract

Two metallo-proteinases of human neutrophil leucocytes, collagenase and gelatinase, were studied. Collagenase specifically cleaved native collagen into the TCA and TCB fragments, whereas gelatinase degraded denatured collagen, i.e. gelatin, and the TCA fragments produced by collagenase. On subcellular fractionation by zonal sedimentation, collagenase was found to be localized in the specific granules, separate from gelatinase, which was recovered in smaller subcellular organelles known as C-particles.Neither enzyme was present in the azurophil granules, which contain the two major serine proteinases of neutrophils, elastase and cathepsin G. Collagenase and gelatinase were separated by gel filtration from extracts of partially purified granules. Both enzymes were found to occur in latent forms and were activated either by trypsin or by 4-aminophenylmercuric acetate. Gelatinase was also activated by cathepsin G, which, however, destroyed collagenase. Both enzymes were destroyed by neutrophil elastase. Activation resulted in a decrease by 25000 in the apparent mol.wt. of both latent metallo-proteinases.

Gelatinase-mediated migration and invasion of cancer cells

“…MMP-2 is abundantly expressed in normal fibroblasts, endothelial and epithelial cells as well as in many transformed cells (Giannelli et al, 1997;Hipps et al, 1991;Partridge et al, 1997;Vartio and Vaheri, 1981). MMP-9 expression is observed in normal leukocytes as well as in transformed cells (Murphy et al, 1980;Sopata and Wize, 1979;Vartio et al, 1982). The genes encoding the gelatinases have been cloned (Huhtala et al, 1990;Huhtala et al, 1991) and these enzymes can be purified with gelatin-affinity chromatography (Hibbs et al, 1985;Johansson and Smedsrod, 1986;Vartio and Vaheri, 1981;Vartio et al, 1982).…”

Proteinases in Human Polymorphonuclear Leukocytes

“…The activity of an enzyme which degrades denatured collagen has previously been reported in preparations of polymorphonuclear leukocytes [16,17]. In one case, it was suggested that the partially purified enzyme degraded triplehelical TCA fragments derived from collagen by digestion with collagenase [17].…”

Purification and characterization of human 92-kDa type IV collagenase (gelatinase B)

“…Considering previous data on two step chromatography techniques for the same enzyme (Hibbs et al 1985), the presently described two step purification method is much simpler and more convenient. The purification of metalloproteinases from human polymorphonuclear neutrophil leukocytes is technically difficult for two main reasons: (a) the metalloproteinase contents are relatively low, and (b) both enzymes are degraded by the serine proteinases, which are present in large amounts in the azurophil granules (Murphy et al, 1980). It has been calculated that 1!10 9 neutrphils contain approx.…”

“…Since its description by Sopata and Dancewicz (Sopata and Dancewicz,1974), human neutrophil gelatinase has been examined by several investigators (Murphy et al, 1980(Murphy et al, , 1982, and Hibbs et al (1985) described 92-kDa type IV collagenase from human neutrophils in 1985.…”