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Dynamic coupling of fast channel gating with slow ATP-turnover underpins protein transport through the Sec translocon

JA. Crossley, WJ. Allen, DW. Watkins, T. Sabir, SE. Radford, R. Tuma, I. Collinson, T. Fessl

. 2024 ; 43 (1) : 1-13. [pub] 20231215

Language English Country England, Great Britain

Document type Journal Article

Persistent link   https://www.medvik.cz/link/bmc24007717

Grant support
BB/V001531/1 UKRI | Biotechnology and Biological Sciences Research Council (BBSRC)
BB/T006889/1 UKRI | Biotechnology and Biological Sciences Research Council (BBSRC)
BB/T008059/1 UKRI | Biotechnology and Biological Sciences Research Council (BBSRC)
RSRP\R1\211057 Royal Society (The Royal Society)
CZ.02.1.01/0.0/0.0/15_003/0000441 EC | European Regional Development Fund (ERDF)
20-11563Y Ministerstvo Vnitra České Republiky (Ministry of the Interior of the Czech Republic)
NA Leeds Beckett University (Leeds Beckett)

The Sec translocon is a highly conserved membrane assembly for polypeptide transport across, or into, lipid bilayers. In bacteria, secretion through the core channel complex-SecYEG in the inner membrane-is powered by the cytosolic ATPase SecA. Here, we use single-molecule fluorescence to interrogate the conformational state of SecYEG throughout the ATP hydrolysis cycle of SecA. We show that the SecYEG channel fluctuations between open and closed states are much faster (~20-fold during translocation) than ATP turnover, and that the nucleotide status of SecA modulates the rates of opening and closure. The SecY variant PrlA4, which exhibits faster transport but unaffected ATPase rates, increases the dwell time in the open state, facilitating pre-protein diffusion through the pore and thereby enhancing translocation efficiency. Thus, rapid SecYEG channel dynamics are allosterically coupled to SecA via modulation of the energy landscape, and play an integral part in protein transport. Loose coupling of ATP-turnover by SecA to the dynamic properties of SecYEG is compatible with a Brownian-rachet mechanism of translocation, rather than strict nucleotide-dependent interconversion between different static states of a power stroke.

References provided by Crossref.org

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$a The Sec translocon is a highly conserved membrane assembly for polypeptide transport across, or into, lipid bilayers. In bacteria, secretion through the core channel complex-SecYEG in the inner membrane-is powered by the cytosolic ATPase SecA. Here, we use single-molecule fluorescence to interrogate the conformational state of SecYEG throughout the ATP hydrolysis cycle of SecA. We show that the SecYEG channel fluctuations between open and closed states are much faster (~20-fold during translocation) than ATP turnover, and that the nucleotide status of SecA modulates the rates of opening and closure. The SecY variant PrlA4, which exhibits faster transport but unaffected ATPase rates, increases the dwell time in the open state, facilitating pre-protein diffusion through the pore and thereby enhancing translocation efficiency. Thus, rapid SecYEG channel dynamics are allosterically coupled to SecA via modulation of the energy landscape, and play an integral part in protein transport. Loose coupling of ATP-turnover by SecA to the dynamic properties of SecYEG is compatible with a Brownian-rachet mechanism of translocation, rather than strict nucleotide-dependent interconversion between different static states of a power stroke.
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