PURIFICATION AND PROPERTIES OF A PROLYL ENDOPEPTIDASE FROM RABBIT BRAIN

被引：203

作者：

ORLOWSKI, M

WILK, E

PEARCE, S

WILK, S

机构：

[1] Department of Pharmacology, Mount Sinai School of Medicine, City University of New York, New York, New York, 10029

来源：

JOURNAL OF NEUROCHEMISTRY | 1979年 / 33卷 / 02期

关键词：

D O I：

10.1111/j.1471-4159.1979.tb05176.x

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Abstract— An enzyme with the specificity of a prolyl endopeptidase was purified about 880‐fold from rabbit brain. The enzyme hydrolyzes peptidylprolyl‐peptide and peptidylprolyl‐amino acid bonds. Several biologically active peptides such as angiotensin, bradykinin, neurotensin. substance P and thyrotropin releasing hormone are degraded by hydrolysis of the bond between the carboxyl group of proline and the adjacent amino acid or ammonia respectively. The enzyme is activated by dithiothreitol and inhibited by heavy metals and thiol blocking agents. The serine protease inhibitor phenylmethanesulfonylfluoride has no effect on activity; however, inhibition was obtained with diisopropylfluorophosphate. Prolyl endopeptidase has a molecular weight of about 66,000 and a pH optimum of about 8.3. A new chromogenic substrate, N‐benzyloxycarbonylglycyl‐L‐prolylsulfamethoxazole, was used for determination of enzyme activity. The substrate is hydrolyzed to N‐benzyloxycarbonylglycyl‐L‐proline and free sulfamethoxazole which can be conveniently determined by a colorimetric procedure. Copyright © 1979, Wiley Blackwell. All rights reserved

引用

页码：461 / 469

页数：9

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