文献类型：期刊文献

英文题名：Chaperone-substrate interactions monitored via a robust TEM-1 β-lactamase fragment complementation assay

作者：Bai, Ling[1];He, Wei[1];Li, Tianpeng[1];Yang, Cuiting[1];Zhuang, Yingping[1];Quan, Shu[1]

机构：[1]East China Univ Sci & Technol, SCICB, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2017

卷号：39

期号：8

起止页码：1191

外文期刊名：BIOTECHNOLOGY LETTERS

收录：;EI(收录号：20171903657675);WOS:【SCI-EXPANDED(收录号:WOS:000405439000011)】；

基金：This study was supported by the National Natural Science Foundation of China (NSFC) (Grant Nos. 31400664 and 3161101097 to S Quan), the Shanghai Pujiang Program (Grant No. 14PJ1402300 to S Quan), and the Fundamental Research Funds for the Central Universities (Grant No. WF1414012 to S Quan).

语种：英文

外文关键词：Cell envelope; Chaperone; Protein fragment complementation assay; Protein-protein interaction; Stress; TEM-1 beta-lactamase

摘要：Objective To investigate the application of the TEM-1 beta-lactamase protein fragment complementation assay (PCA) in detecting weak and unstable protein-protein interactions as typically observed during chaperone-assisted protein folding in the periplasm of Escherichia coli. Results The TEM-1 beta-lactamase PCA system effectively captured the interactions of three pairs of chaperones and substrates. Moreover, the strength of the interactions can be quantitatively analyzed by comparing different levels of penicillin resistance, and the assay can be performed under 0.5% butanol, a stress condition thought to be physiologically relevant. Conclusions The beta-lactamase PCA system faithfully reports chaperone-substrate interactions in the bacterial cell envelope, and therefore this system has the potential to map the complex protein homeostasis network under a fluctuating environment.