Chaperone-substrate interactions monitored via a robust TEM-1 beta-lactamase fragment complementation assay
Chaperone-substrate interactions monitored via a robust TEM-1 beta-lactamase fragment complementation assay
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通过强大的 TEM-1 β-内酰胺酶片段互补测定法监测分子伴侣 - 底物相互作用
DOI：
10.1007/s10529-017-2347-9
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发表时间：
2017
影响因子：
2.7
通讯作者：
Quan Shu
中科院分区：
文献类型：
--
作者：
Bai Ling;He Wei;Li Tianpeng;Yang Cuiting;Zhuang Yingping;Quan Shu
目的探讨TEM-1 β-内酰胺酶蛋白片段互补分析（PCA）在检测大肠杆菌周质中伴侣辅助蛋白折叠过程中常见的弱且不稳定的蛋白质-蛋白质相互作用中的应用。结果TEM-1 β-内酰胺酶PCA系统有效捕获了三对伴侣和底物的相互作用。此外，可以通过比较不同水平的青霉素耐药性来定量分析相互作用的强度，并且可以在0.5%丁醇（一种被认为具有生理相关性的应激条件）下进行测定。结论β-内酰胺酶PCA系统忠实地报告了细菌细胞包膜中的分子伴侣-底物相互作用，因此该系统具有绘制波动环境下复杂蛋白质稳态网络的潜力。
ObjectiveTo investigate the application of the TEM-1 β-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 ofEscherichia coli.ResultsThe TEM-1 β-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.ConclusionsThe β-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.