用于测定丁酰胆碱酯酶和芳基酯酶活性的新型显色和荧光底物。

New chromogenic and fluorogenic substrates for the determination of butyrylcholinesterase and arylesterase activities.

作者信息

Nozawa M, Tanizawa K, Kanaoka Y

出版信息

J Pharmacobiodyn. 1980 Jul;3(7):321-7. doi: 10.1248/bpb1978.3.321.

DOI:10.1248/bpb1978.3.321

PMID:7205543

Abstract

A simple direct spectrophotometric method for the determination of butyrylcholinesterase (EC 3.1.1.8) and arylesterase (EC 3.1.1.2) activities has been developed. New chromogenic substrates, (3-carboxypropyl)trimethylammonium iodide o-nitrophenyl ester (I) and (3-carboxypropyl)trimethylammonium iodide p-nitrophenyl ester (II), as well as new fluorogenic substrate, (3-carboxypropyl)trimethylammonium iodide 4'-methylumbelliferyl ester (III), were used in this study. Horse serum butyrylcholinesterase equally catalyzed hydrolysis of the compounds, I, II and III. Hydrolysis of these compounds by trypsin, chymotrypsin, acetylcholinesterase and carboxylesterase was negligible or quite slow. By human serum butyrylcholinesterase, however, only the compound I was preferentially hydrolyzed. The compound III, by contrast, was found to be a specific substrate for arylesterase of human serum without being affected by the butyrylcholinesterase. All these measurements were carried out readily and efficiently, by analyzing highly colored products with I and II, and highly fluorescent product with III.

摘要

已开发出一种用于测定丁酰胆碱酯酶（EC 3.1.1.8）和芳基酯酶（EC 3.1.1.2）活性的简单直接分光光度法。本研究使用了新型显色底物，即（3-羧丙基）三甲基碘化铵邻硝基苯酯（I）和（3-羧丙基）三甲基碘化铵对硝基苯酯（II），以及新型荧光底物，即（3-羧丙基）三甲基碘化铵4'-甲基伞形酮酯（III）。马血清丁酰胆碱酯酶能同等程度地催化化合物I、II和III的水解。胰蛋白酶、胰凝乳蛋白酶、乙酰胆碱酯酶和羧酸酯酶对这些化合物的水解作用可忽略不计或相当缓慢。然而，人血清丁酰胆碱酯酶仅优先水解化合物I。相比之下，发现化合物III是人类血清芳基酯酶的特异性底物，不受丁酰胆碱酯酶的影响。通过分析I和II产生的高显色产物以及III产生的高荧光产物，所有这些测量都能轻松高效地进行。