Inhibition of membrane-type serine protease 1/matriptase by natural and synthetic protease inhibitors.
Membrane-type serine protease 1 (MT-SP1), identical to matriptase, is a recently identified type II transmembrane serine protease. MT-SP1/matriptase is of considerable interest for the development, homeostasis, and cancer invasion and metastasis of epithelial tissues. The administration of inhibitors for MT-SP1/matriptase may be effective to suppress the development of tumors where the enzyme may be involved. In the present study, we produced a secreted form of recombinant MT-SP1/matriptase (ekMT-SP1s) that can be activated by enterokinase in vitro and investigated the inhibitory ability of various protease inhibitors toward the recombinant enzyme. The enterokinase-treated ekMT-SP1s (active ekMT-SP1s) cleaved various peptidyl-4-methylcoumaryl-7-amide (MCA) substrates with arginine (or lysine) residue at position P1, and the best substrate was t-butyloxycarbonyl (Boc)-Gln-Ala-Arg-MCA. The specificity for the synthetic and natural substrates of the active ekMT-SP1s was in good agreement with that of the natural enzyme. Endogenous protease inhibitors tested, except for antithrombin III, showed no or little inhibition on the cleavage of Boc-Gln-Ala-Arg-MCA by the active ekMT-SP1s. Aprotinin showed strong inhibitory activity toward the cleavage. Food-derived inhibitors, such as soybean trypsin inhibitor, Bowman-Birk inhibitor, and lima bean trypsin inhibitor inhibited it, while chicken ovomucoid did not. Synthetic inhibitors tested inhibited it, and among them, the inhibitory effect of FOY-305 was strongest. The present findings provide important information for the suppression of cancer invasion and metastasis for which MT-SP1/matriptase is responsible.
Yamasaki
Yoshie
Y
Laboratory of Nutrition Chemistry, Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan.
Satomi
Shigeki
S
Murai
Nobuhito
N
Tsuzuki
Satoshi
S
Fushiki
Tohru
T
D000818:Animals; D000990:Antithrombin III / Q000494:pharmacology; D007611:Aprotinin / Q000494:pharmacology; D019556:COS Cells; D004591:Electrophoresis, Polyacrylamide Gel; D004765:Enteropeptidase / Q000378:metabolism; D006801:Humans; D010059:Ovomucin / Q000494:pharmacology; D010940:Plant Proteins / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology*; D011994:Recombinant Proteins; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D013025:Glycine max / Q000737:chemistry; D013379:Substrate Specificity; D014162:Transfection; D014357:Trypsin / Q000235:genetics / Q000378:metabolism*; D014358:Trypsin Inhibitor, Bowman-Birk Soybean / Q000494:pharmacology; D014359:Trypsin Inhibitor, Kazal Pancreatic / Q000494:pharmacology; D000515:alpha 1-Antitrypsin / Q000494:pharmacology; D000511:alpha-Macroglobulins / Q000494:pharmacology
D010940:Plant Proteins; D011480:Protease Inhibitors; D011994:Recombinant Proteins; D014358:Trypsin Inhibitor, Bowman-Birk Soybean; D000515:alpha 1-Antitrypsin; D000511:alpha-Macroglobulins; C025398:protease inhibitor protein, Phaseolus lunatus; D010059:Ovomucin; D014359:Trypsin Inhibitor, Kazal Pancreatic; D000990:Antithrombin III; D007611:Aprotinin; D012697:Serine Endopeptidases; C485595:matriptase; C485596:ST14 protein, human; D014357:Trypsin; D004765:Enteropeptidase