SUICIDE-SUBSTRATE INACTIVATION OF BETA-GALACTOSIDASE BY DIAZOMETHYL BETA-D-GALACTOPYRANOSYL KETONE

被引:6
作者
BEMILLER, JN [1 ]
GILSON, RJ [1 ]
MYERS, RW [1 ]
SANTORO, MM [1 ]
机构
[1] SO ILLINOIS UNIV, DEPT CHEM & BIOCHEM, CARBONDALE, IL 62901 USA
基金
美国国家卫生研究院;
关键词
D O I
10.1016/0008-6215(93)84159-4
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
Diazomethyl beta-D-galactopyranosyl ketone (1) has been proven to be a mechanism-based, irreversible (suicide-substrate) inactivator of Aspergillus oryzae beta-D-galactosidase, but not an inactivator of E. coli lacZ beta-D-galactosidase, Compound 1 is stable in buffers of normal physiological pH. It is decomposed by H+, but not by nucleophiles. Inactivation of A. oryzae beta-D-galactosidase was proven to be mechanism-based and irreversible via experiments which showed that the enzyme could be protected from inactivation by a competitive inhibitor, neither diazomethyl beta-D-glucopyranosyl ketone (2) nor diazomethyl alpha-D-galactopyranosyl ketone inactivated the enzyme and therefore inactivation is stereospecific, excess inhibitor could be separated from inactive enzyme without regain of activity and therefore it is bound irreversibly, and a second pulse of enzyme is inactivated at the same rate as enzyme inactivated to 95% activity by the first pulse. Diazomethyl beta-D-glucopyranosyl ketone (2) inhibited sweet almond beta-D-glucosidase.
引用
收藏
页码:101 / 112
页数:12
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