원문정보
초록
영어
Metalloproteases are related to various diseases such as bacterial infections, cholera, gastritis and peptic ulcer, and gastric carcinoma. Hence sensitive and selective assay materials of metalloproteases activity need to be developed for the detection of those diseases. In this study, we synthesized five DBDY-(amino acid-INH)2 from the conjugation of N,N′-diBoc-dityrosine (DBDY) with two molecules of amino acid and isoniazid (INH). The hydrolysis reaction resulted in the release of Amino acid-INH, and recovered fluorescence of DBDY. These five DBDY-(amino acid-INH)2 were tested for hydrolysis by four metalloproteases; thermolysin, dispase, collagenase, and carboxypeptidase A. As a result, DBDY-(Phe-INH)2 and DBDY-(Ile-INH)2 were found to be the substrates for the sensitive and selective assay of thermolysin and dispase. The sensitivity of thermolysin to DBDY-(Phe-INH)2 was evaluated at various concentrations of thermolysin while the initial concentration of DBDY- (Phe-INH)2 was fixed at 2.5 μM. DBDY-(Phe-INH)2 allowed the detectable reaction with thermolysin up to 1 pM and showed the apparent increase in the fluorescence. Therefore, DBDY-(Phe-INH)2 and DBDY-(Ile-INH)2 are believed to be useful for the sensitive and selective assay of metalloproteases such as thermolysin and dispase.