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Bacterial Hsp70 resolves misfolded states and accelerates productive folding of a multi-domain protein.
Nature Communications ( IF 14.7 ) Pub Date : 2020-01-17 , DOI: 10.1038/s41467-019-14245-4
Rahmi Imamoglu 1 , David Balchin 1 , Manajit Hayer-Hartl 1 , F Ulrich Hartl 1
Affiliation  

The ATP-dependent Hsp70 chaperones (DnaK in E. coli) mediate protein folding in cooperation with J proteins and nucleotide exchange factors (E. coli DnaJ and GrpE, respectively). The Hsp70 system prevents protein aggregation and increases folding yields. Whether it also enhances the rate of folding remains unclear. Here we show that DnaK/DnaJ/GrpE accelerate the folding of the multi-domain protein firefly luciferase (FLuc) ~20-fold over the rate of spontaneous folding measured in the absence of aggregation. Analysis by single-pair FRET and hydrogen/deuterium exchange identified inter-domain misfolding as the cause of slow folding. DnaK binding expands the misfolded region and thereby resolves the kinetically-trapped intermediates, with folding occurring upon GrpE-mediated release. In each round of release DnaK commits a fraction of FLuc to fast folding, circumventing misfolding. We suggest that by resolving misfolding and accelerating productive folding, the bacterial Hsp70 system can maintain proteins in their native states under otherwise denaturing stress conditions.

中文翻译:

细菌热休克蛋白70解析错误折叠状态,并加速多域蛋白的生产性折叠。

ATP依赖的Hsp70分子伴侣(在大肠杆菌中为DnaK)与J蛋白和核苷酸交换因子(分别为大肠杆菌DnaJ和GrpE)协作介导蛋白质折叠。Hsp70系统可防止蛋白质聚集并提高折叠产量。能否提高折叠速度还不清楚。在这里,我们显示DnaK / DnaJ / GrpE加速了多域蛋白萤火虫萤光素荧光素酶(FLuc)的折叠,其速度是在没有聚集的情况下测得的自发折叠速率的20倍。通过单对FRET和氢/氘交换的分析确定域间错折叠是缓慢折叠的原因。DnaK结合扩大了错误折叠的区域,从而解决了动力学捕获的中间体,折叠发生在GrpE介导的释放中。在每轮发布中,DnaK都会将FLuc的一部分投入快速折叠,从而避免错误折叠。我们建议通过解决错误折叠和加快生产性折叠,细菌Hsp70系统可以在其他条件下使蛋白质处于变性状态,从而保持应激状态。
更新日期:2020-01-17
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