Two electrical potential-dependent steps are required for transport by the Escherichia coli Tat machinery.

Two electrical potential-dependent steps are required for transport by the Escherichia coli Tat machinery.
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通过大肠杆菌tat机械运输需要两个电势依赖性步骤。

DOI:
10.1083/jcb.200702082
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发表时间:
2007-10-08
影响因子:
7.8
通讯作者:
Musser, Siegfried M.
Musser, Siegfried M.
中科院分区:
生物学1区
文献类型:
--
作者:
Bageshwar, Umesh K.;Musser, Siegfried M.

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大肠杆菌中的双精氨酸转运(达特)途径转运完全折叠和组装的蛋白质穿过能量转导周质膜。在叶绿体中,达特运输只需要来自质子动力的能量输入。为了阐明细菌达特蛋白转运的机制和能量学,我们开发了一种有效的体外转运试验,使用富含TatABC的倒置膜囊泡和生理前体前SufI。我们报告的运输效率为60-80%纳摩尔前SufI浓度。pH梯度的消散不会降低前SufI转运效率。相反,前SufI转运需要至少两个依赖于电位(Δ V)的步骤,其在所需Δ V的持续时间和最小幅度上都不同。这些数据与一个模型是一致的,在该模型中,早期的传输步骤需要大量的短持续时间的Δψ,而在该模型中,驱动后期的传输步骤需要一个小的长持续时间的Δψ。
The twin-arginine translocation (Tat) pathway in Escherichia coli transports fully folded and assembled proteins across the energy-transducing periplasmic membrane. In chloroplasts, Tat transport requires energy input only from the proton motive force. To elucidate the mechanism and energetics of bacterial Tat protein transport, we developed an efficient in vitro transport assay using TatABC-enriched inverted membrane vesicles and the physiological precursor pre-SufI. We report transport efficiencies of 60–80% for nanomolar pre-SufI concentrations. Dissipation of the pH gradient does not reduce pre-SufI transport efficiency. Instead, pre-SufI transport requires at least two electrical potential (Δψ)–dependent steps that differ in both the duration and minimum magnitude of the required Δψ. The data are consistent with a model in which a substantial Δψ of short duration is required for an early transport step, and in which a small Δψ of long duration is necessary to drive a later transport step.
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