Substrate specificity of a novel alcohol resistant metalloproteinase, vimelysin, from Vibrio sp. T1800.
Vimelysin is a novel alcohol resistant metalloproteinase from Vibrio sp. T1800. The substrate specificity of vimelysin was studied by using natural and furylacryloyl dipeptide substrates. Vimelysin cleaved mainly Pro7-Phe8 bond and slightly Tyr4-Ile5 bond in human angiotensin I. Vimelysin also cleaved mainly Phe24-Phe25 and Tyr16-Leu17 bonds, and slightly His5-Leu6, His10-Leu11, Ala14-Leu15, and Gly23-Phe24 bonds in oxidized insulin B-chain. The substrate specificity of vimelysin, by using furylacryloyl (Fua) dipeptides were also studied. The ratio of kcat/Km for Fua-Gly-Phe-NH2/Fua-Gly-Leu-NH2, Fua-Phe-Leu-NH2/Fua-Gly-Leu-NH2, and Fua-Phe-Phe-NH2/Fua-Gly-Leu-NH2 were 15.9, 27.8, and 59.0, respectively. These results indicate that vimelysin easily recognizes phenylalanine in P1' positions, which is different from thermolysin.
Takahashi
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Department of Applied Biology, Faculty of Textile Science, Kyoto Institute of Technology, Japan.
Okayama
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Kunugi
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Oda
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K
D000595:Amino Acid Sequence; D000803:Angiotensin I / Q000737:chemistry; D001426:Bacterial Proteins / Q000378:metabolism*; D004151:Dipeptides / Q000378:metabolism*; D004352:Drug Resistance, Microbial; D000431:Ethanol / Q000494:pharmacology*; D005456:Fluorescent Dyes; D007328:Insulin / Q000737:chemistry; D007700:Kinetics; D008666:Metalloendopeptidases / Q000378:metabolism*; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000737:chemistry; D013379:Substrate Specificity; D014733:Vibrio / Q000201:enzymology*
D001426:Bacterial Proteins; D004151:Dipeptides; D005456:Fluorescent Dyes; D007328:Insulin; D010446:Peptide Fragments; D000431:Ethanol; D000803:Angiotensin I; D008666:Metalloendopeptidases; C099068:vimelysin