Substrate-triggered position switching of TatA and TatB during Tat transport in Escherichia coli.

Substrate-triggered position switching of TatA and TatB during Tat transport in Escherichia coli.
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大肠杆菌中TAT运输过程中TATA和TATB的底物触发位置切换。

DOI:
10.1098/rsob.170091
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发表时间:
2017-08
期刊:
影响因子:
5.8
通讯作者:
Palmer T
Palmer T
中科院分区:
生物学2区
文献类型:
--
作者:
Habersetzer J;Moore K;Cherry J;Buchanan G;Stansfeld PJ;Palmer T

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双精氨酸蛋白转运(达特)机制介导折叠蛋白质跨原核生物细胞质膜和植物叶绿体类囊体膜的转运。大肠杆菌达特系统包含TatC和两种另外的序列相关蛋白质TatA和TatB。活性易位酶按需组装,底物结合在TatABC受体复合物上触发TatA的多个额外拷贝的募集和组装;然而,介导易位酶组装的分子相互作用知之甚少。TatC跨膜(TM)螺旋5上的“极性簇”位点先前被鉴定为与TatB结合。在这里,我们使用二硫键交联和分子建模,以确定一个新的结合位点上的TatC TM螺旋6,相邻的极性集群网站。我们证明,TatA和TatB每个都有能力结合在两个TatC网站,但在体内,这是根据复杂的激活状态进行调节。在静息态系统中,TatB结合极性簇位点,TatA占据TM螺旋6位点。然而,当系统被激活的过度生产的底物,TatA和TatB开关结合位点。我们建议,这种基板触发的位置交换是一个活跃的达特易位酶的组装中的关键步骤。
The twin-arginine protein transport (Tat) machinery mediates the translocation of folded proteins across the cytoplasmic membrane of prokaryotes and the thylakoid membrane of plant chloroplasts. The Escherichia coli Tat system comprises TatC and two additional sequence-related proteins, TatA and TatB. The active translocase is assembled on demand, with substrate-binding at a TatABC receptor complex triggering recruitment and assembly of multiple additional copies of TatA; however, the molecular interactions mediating translocase assembly are poorly understood. A ‘polar cluster’ site on TatC transmembrane (TM) helix 5 was previously identified as binding to TatB. Here, we use disulfide cross-linking and molecular modelling to identify a new binding site on TatC TM helix 6, adjacent to the polar cluster site. We demonstrate that TatA and TatB each have the capacity to bind at both TatC sites, however in vivo this is regulated according to the activation state of the complex. In the resting-state system, TatB binds the polar cluster site, with TatA occupying the TM helix 6 site. However when the system is activated by overproduction of a substrate, TatA and TatB switch binding sites. We propose that this substrate-triggered positional exchange is a key step in the assembly of an active Tat translocase.
DOI: 10.1021/ct300646g
发表时间: 2013-01-01
影响因子: 5.5
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