An in vivo protease activity assay for investigating the functions of the Escherichia coli membrane protease HtpX
An in vivo protease activity assay for investigating the functions of the Escherichia coli membrane protease HtpX
复制标题
用于研究大肠杆菌膜蛋白酶 HtpX 功能的体内蛋白酶活性测定
DOI：
10.1002/1873-3468.13368
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发表时间：
2019
期刊：
影响因子：
3.5
通讯作者：
Akiyama Yoshinori
中科院分区：
文献类型：
--
作者：
Yoshitani Kohei;Hizukuri Yohei;Akiyama Yoshinori
大肠埃希菌HtpX是一种位于细胞膜上的M48家族锌金属蛋白酶。以往的研究表明，它参与了膜蛋白的质量控制。然而，它的活体蛋白分解功能还没有得到详细的表征，主要是因为生理底物还没有确定，也没有能够灵敏地检测蛋白酶活性的模型底物。我们构建了一种新的HtpX模型底物，并建立了一种活的、定量的、简便的HtpX蛋白酶活性测定体系。该系统能够检测在保守区域携带突变的HtpX突变体的差异蛋白水解酶活性。该系统也可用于研究HtpX及其同源物在其他细菌中的功能。
Escherichia coliHtpX is an M48 family zinc metalloproteinase located in the cytoplasmic membrane. Previous studies suggested that it is involved in the quality control of membrane proteins. However, itsin vivoproteolytic function has not been characterized in detail, mainly because the physiological substrates have not been identified and no model substrate that allows sensitive detection of the protease activity is available. We constructed a new model substrate of HtpX and established anin vivosemiquantitative and convenient protease activity assay system for HtpX. This system enables detection of differential protease activities of HtpX mutants carrying mutations in conserved regions. This system would also be useful for investigating the functions of HtpX and its homologs in other bacteria.