S-Hippuryl Thioglycolyl Glycine : A New Chromogenic Substrate for Dipeptidyl Carboxypeptidases

국문 초록 (Abstract)

Dideptidyl carboxypeptidases와 angiotenisn-coverting enzyme의 새로운 기질불질로서 thiol ester 인 S-Hippuryl thioglycolyl glycine을 합성하였으며, 이 기질에 의한 간편하고도 예민한 효소 활성도의 정량방법을 제시하였다. 이 경우 효소반응 생성물인 thioglycolyl glycine은 반응계중에 첨가한 5,5-dithio-bis-(2-nitrobenzoic acid), DTNB와 쉽게 반응하여 410nm에서 강한 흡광스펙트럼을 갖는 5-thio-2-nitrobenzoic acid(${\varepsilon}M=1.36{\times}10^4$)을 형성함으로서 효소의 새로운 미량정량 방법으로 이용 가치가 크다고 본다.

다국어 초록 (Multilingual Abstract)

A spectrophotometric assay technique is descriead for the measurement of free SH-groups in the enzyme reaction mixture. The method utilizes a new substrate, S-hippuryl-thioglycolyl-glycine(S-Hip-thioglycol-Gly) which is the basis for a convenient assay of angiotensin-converting enzyme and other dipeptidyl carboxypeptidases. This substrate contains an appropriately located thioester linkage that is hydrolyzed by the converting enzyme and other dipeptidyl carboxypeptidases. One of the products, thioglycolyl glycine, is readily measured by reaction with Ellman's reagent, 5,5'-dithio-bis-(2-nitrobenzoic acid), DTNB, to produce 5-thio-2-nitrobenzoic acid which has a strong absorption band at 410 nm. The method is sensitive (${\varepsilon}M = 1.36{\times}10^4$ at 412 nm) and can be applied as a continuous recording with DTNB present in the enzymatic reaction mixture.