一种在存在蒽黄素的情况下用丙氨酸脱氢酶测定γ-谷氨酰环转移酶的分光光度法。

A spectrophotometric method for the determination of gamma-glutamyl cyclotransferase with alanine dehydrogenase in the presence of anthglutin.

作者信息

Takahashi T, Kondo T, Ohno H, Minato S, Ohshima T, Mikuni S, Soda K, Taniguchi N

机构信息

Department of Biochemistry, Osaka University Medical School, Japan.

出版信息

Biochem Med Metab Biol. 1987 Dec;38(3):311-6. doi: 10.1016/0885-4505(87)90095-8.

DOI:10.1016/0885-4505(87)90095-8

PMID:2893631

Abstract

gamma-Glutamyl cyclotransferase activity is assayed in tissues by a colorimetric method using gamma-glutamyl alanine as a substrate coupled with alanine dehydrogenase from B. sphericus, to measure the formation of NADH. In order to avoid interference by the reaction catalyzed by gamma-glutamyl transpeptidase, anthglutin, a specific inhibitor of the transpeptidase was included in the reaction mixture. The Km value of rat kidney gamma-glutamyl cyclotransferase with respect to gamma-glutamyl alanine appeared to be the same when determined by either the colorimetric or the radiometric method. This assay presents a reliable alternative to the use of radiolabeled substrate and is used for the assay of gamma-glutamyl cyclotransferase in a variety of physiological and experimental samples.

摘要

采用比色法，以γ-谷氨酰丙氨酸为底物，与球形芽孢杆菌的丙氨酸脱氢酶偶联，通过测定NADH的生成量来检测组织中的γ-谷氨酰环化转移酶活性。为避免γ-谷氨酰转肽酶催化的反应产生干扰，反应混合物中加入了转肽酶的特异性抑制剂氨硫脲。用比色法或放射性测定法测定时，大鼠肾脏γ-谷氨酰环化转移酶对γ-谷氨酰丙氨酸的Km值似乎相同。该检测方法是使用放射性标记底物的可靠替代方法，可用于检测各种生理和实验样品中的γ-谷氨酰环化转移酶。