A cell-based assay that targets methionine aminopeptidase in a physiologically relevant environment

被引：8

作者：

Chai, Sergio C. ^{[1
]}

Ye, Qi-Zhuang ^{[1
]}

机构：

[1] Indiana Univ, Sch Med, Dept Biochem & Mol Biol, Indianapolis, IN 46202 USA

来源：

BIOORGANIC & MEDICINAL CHEMISTRY LETTERS | 2010年 / 20卷 / 07期

关键词：

Cell-based assay; Metalloenzyme; Enzyme inhibitor; Antibacterial; Drug screening; ESCHERICHIA-COLI; SACCHAROMYCES-CEREVISIAE; INHIBITORS; DISCOVERY; GENE;

D O I：

10.1016/j.bmcl.2010.02.052

中图分类号：

R914 [药物化学];

学科分类号：

100701 ;

摘要：

Methionine aminopeptidase (MetAP) is a promising target for the development of novel antibiotics. However, many potent inhibitors of the purified enzyme failed to show significant antibacterial activity. It is uncertain which divalent metal MetAP uses as its native cofactor in bacterial cells. Herein, we describe a cell-based assay that monitors the hydrolysis of a fluorogenic substrate by overexpressed MetAP in permeabilized Escherichia coli cells and its validation with a set of MetAP inhibitors. This cell-based assay is applicable to those cellular targets with poorly defined native cofactor, increasing the chances of identifying inhibitors that can inhibit the cellular target. (C) 2010 Elsevier Ltd. All rights reserved.

引用

页码：2129 / 2132

页数：4

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