| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1991: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1990: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Research Abstract |
Proteolytic enzymes degrading extracellular matrix and basement membrane are thought to play important roles in tumor metastasis. High activity of type IV collagenase is correlated to the metastatic potential of tumor.
A new protease degrading type IV collagen was purified 8, 000-folds from human stomach carcinoma and 1, 200 folds from metastatic hepatic tumor. The enzyme cleave type IV collagen but not type I, II, III, V collagen, fibronectin, casein, albumin or hemoglobin. The optimum pH of the enzyme is about 7.6.
The molecular weight of the enzyme was estimated more than 2, 000 kDa by Sephacryl S300 and Superose 6HR gel filtration. It is higher than the other established type IV collagenase, for example MMP (Matrix metalloprotease) -2 (m. w. 72 kDa) or MMP-9 (m. w. 92kDa). The purified enzyme was inactive forni, howevet it showed collagenolitic activity by trypsin-treatment. APMA (Paminophenyulmercuric acetate) -treatment did not activate the inactive enzyme, though it is an activator of latent form metalloprotease. The enzyme is inhibited by metalloproteaseinhibitors but not by TIMP (Tissue inhibitor of metalloprotease). These properties of the new enzyme are different from the other type IV collagenase reported.
Now, to proceed this research, specific antibody of the new enzyme has jest isolated from rabbit anti-new enzyme sera.
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