Activation of the precursor of human stromelysin 2 and its interactions with other matrix metalloproteinases.
Matrix metalloproteinases (MMP) are synthesized as inactive zymogens (proMMP) and subsequently activated by many factors to degrade the extracellular matrix (ECM). In the present study, we have examined the intermolecular activation mechanisms of proMMP by MMP-10 (stromelysin 2). ProMMP-10 was purified from the culture media of OSC-20 human oral squamous carcinoma cells stimulated with 12-O-tetradecanoylphorbol 13-acetate. The final products are partially activated (approximately 38% of the full activity) during the purification steps and contain proMMP-10 of Mr 56,000 with minor protein bands of Mr 47,000, 24,000 and 22,000. The zymogen is activated by 4-aminophenylmercuric acetate and processed to the active forms of Mr 47,000 and 24,000. The NH2-terminal sequence of the 47,000- and 24,000-Mr species is Phe82-Ser-Ser-Phe-Pro-Gly, which is identical to that of stromelysin 2. ProMMP-9 (progelatinase B) is activated by MMP-10 to its full activity and processed to the low-Mr species of Mr 81,000, 65,000, 57,000 and 55,000, the former two of which show proteolytic activity on a gelatin zymography. The NH2-terminal sequence analysis indicates that the 81,000-, 65,000- and 57,000-M, species have the identical sequence of Phe88-Gln-Thr-Phe-Glu-Gly, suggesting the cleavage of the Arg87-Phe88 peptide bond for activation and both NH2-terminal and COOH-terminal truncation in the 65,000- and 57,000-Mr forms. MMP-10 also activates proMMP-7 (promatrilysin) up to about 60% of the full activity and generates the same active species of Mr 19,000 as that obtained by activation with 4-aminophenylmercuric acetate. Incubation of proMMP-2 (progelatinase A) or proMMP-3 with MMP-10 does not result in activation of these proMMP. These results indicate that in addition to the previously reported activation of proMMP-1 (tissue procollagenase) and proMMP-8 (neutrophil procollagenase), MMP-10 can also activate proMMP-9 and proMMP-7, and suggest the possibility that MMP-10 may replace a role of MMP-3 in the ECM degradation in concert with other MMP under various pathological conditions.
Nakamura
H
H
Department of Molecular Immunology and Pathology, Cancer Research Institute, Kanazawa University, Japan.
Fujii
Y
Y
Ohuchi
E
E
Yamamoto
E
E
Okada
Y
Y
D000595:Amino Acid Sequence; D017364:Collagenases / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D004792:Enzyme Precursors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005109:Extracellular Matrix / Q000201:enzymology; D018093:Gelatinases / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D066298:In Vitro Techniques; D053506:Matrix Metalloproteinase 10; D020778:Matrix Metalloproteinase 2; D019278:Matrix Metalloproteinase 3 / Q000378:metabolism; D020783:Matrix Metalloproteinase 7; D020780:Matrix Metalloproteinase 9; D008666:Metalloendopeptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010662:Phenylmercuric Acetate / Q000031:analogs & derivatives / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D004792:Enzyme Precursors; C010024:4-aminophenylmercuriacetate; D017364:Collagenases; D018093:Gelatinases; D008666:Metalloendopeptidases; D019278:Matrix Metalloproteinase 3; D053506:Matrix Metalloproteinase 10; D020783:Matrix Metalloproteinase 7; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D010662:Phenylmercuric Acetate