HDX-MS Reveals Nucleotide-dependent, Anti-correlated Opening and Closure of SecA and SecY Channels of the Bacterial Translocon

Overview

Journal Elife

Specialty Biology

Date 2019 Jul 11

PMID 31290743

Citations 8

Authors

Zainab Ahdash

Euan Pyle

William John Allen

Robin A Corey

Ian Collinson

Argyris Politis

Affiliations

Soon will be listed here.

Abstract

The bacterial Sec translocon is a multi-protein complex responsible for translocating diverse proteins across the plasma membrane. For post-translational protein translocation, the Sec-channel - SecYEG - associates with the motor protein SecA to mediate the ATP-dependent transport of pre-proteins across the membrane. Previously, a diffusional-based Brownian ratchet mechanism for protein secretion has been proposed; the structural dynamics required to facilitate this mechanism remain unknown. Here, we employ hydrogen-deuterium exchange mass spectrometry (HDX-MS) to reveal striking nucleotide-dependent conformational changes in the Sec protein-channel from . In addition to the ATP-dependent opening of SecY, reported previously, we observe a counteracting, and ATP-dependent, constriction of SecA around the pre-protein. ATP binding causes SecY to open and SecA to close; while, ADP produced by hydrolysis, has the opposite effect. This alternating behaviour could help impose the directionality of the Brownian ratchet for protein transport through the Sec machinery.

Citing Articles

AFM observation of protein translocation mediated by one unit of SecYEG-SecA complex.

Kanaoka Y, Mori T, Nagaike W, Itaya S, Nonaka Y, Kohga H Nat Commun. 2025; 16(1):225.

PMID: 39779699 PMC: 11711467. DOI: 10.1038/s41467-024-54875-x.

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

Crossley J, Allen W, Watkins D, Sabir T, Radford S, Tuma R EMBO J. 2024; 43(1):1-13.

PMID: 38177311 PMC: 10883268. DOI: 10.1038/s44318-023-00004-1.

Focusing on Infections: Distribution, Antimicrobial Susceptibilities and Phylogeny.

Pino-Rosa S, Medina-Pascual M, Carrasco G, Garrido N, Villalon P, Valiente M Antibiotics (Basel). 2023; 12(11).

PMID: 37998770 PMC: 10668661. DOI: 10.3390/antibiotics12111568.

A unifying mechanism for protein transport through the core bacterial Sec machinery.

Allen W, Collinson I Open Biol. 2023; 13(8):230166.

PMID: 37643640 PMC: 10465204. DOI: 10.1098/rsob.230166.

Rate-limiting transport of positively charged arginine residues through the Sec-machinery is integral to the mechanism of protein secretion.

Allen W, Corey R, Watkins D, Oliveira A, Hards K, Cook G Elife. 2022; 11.

PMID: 35486093 PMC: 9110029. DOI: 10.7554/eLife.77586.

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