Inter-domain dynamics in the chaperone SurA and multi-site binding to its outer membrane protein clients

Abstract

The periplasmic chaperone SurA plays a key role in outer membrane protein (OMP) biogenesis. E. coli SurA comprises a core domain and two peptidylprolyl isomerase domains (P1 and P2), but its mechanisms of client binding and chaperone function have remained unclear. Here, we use chemical cross-linking, hydrogen-deuterium exchange mass spectrometry, single-molecule FRET and molecular dynamics simulations to map the client binding site(s) on SurA and interrogate the role of conformational dynamics in OMP recognition. We demonstrate that SurA samples an array of conformations in solution in which P2 primarily lies closer to the core/P1 domains than suggested in the SurA crystal structure. OMP binding sites are located primarily in the core domain, and OMP binding results in conformational changes between the core/P1 domains. Together, the results suggest that unfolded OMP substrates bind in a cradle formed between the SurA domains, with structural flexibility between domains assisting OMP recognition, binding and release.

Metadata

Item Type:	Article
Authors/Creators:	Calabrese, AN https://orcid.org/0000-0003-2437-7761 Schiffrin, B https://orcid.org/0000-0002-1670-7356 Watson, M https://orcid.org/0000-0003-2166-6840 Karamanos, TK https://orcid.org/0000-0003-2297-540X Walko, M https://orcid.org/0000-0002-7160-6136 Humes, JR https://orcid.org/0000-0002-1407-7880 Horne, JE https://orcid.org/0000-0001-5260-2634 White, P https://orcid.org/0000-0001-7536-4552 Wilson, AJ https://orcid.org/0000-0001-9852-6366 Kalli, AC https://orcid.org/0000-0001-7156-9403 Tuma, R https://orcid.org/0000-0003-0047-0013 Ashcroft, AE https://orcid.org/0000-0002-1676-123X Brockwell, DJ https://orcid.org/0000-0002-0802-5937 Radford, SE https://orcid.org/0000-0002-3079-8039
Copyright, Publisher and Additional Information:	© The Author(s) 2020. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/ licenses/by/4.0/.
Dates:	Accepted: 18 March 2020 Published (online): 1 May 2020 Published: 1 May 2020
Institution:	The University of Leeds
Academic Units:	The University of Leeds > Faculty of Engineering & Physical Sciences (Leeds) > School of Chemistry (Leeds) > Organic Chemistry (Leeds) The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) > Biomolecular Mass Spectroscopy (Leeds) The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) > Structural Molecular Biology 2 (Leeds) The University of Leeds > Faculty of Medicine and Health (Leeds) > School of Medicine (Leeds) > Leeds Institute of Cardiovascular and Metabolic Medicine (LICAMM) > Discovery & Translational Science Dept (Leeds)
Funding Information:	Funder Grant number BBSRC (Biotechnology & Biological Sciences Research Council) BB/P000037/1 BBSRC (Biotechnology & Biological Sciences Research Council) BB/N007603/1 BBSRC (Biotechnology & Biological Sciences Research Council) BB/T000635/1 BBSRC (Biotechnology & Biological Sciences Research Council) BB/N017307/1 EPSRC (Engineering and Physical Sciences Research Council) EP/N035267/1 MRC (Medical Research Council) MR/P018491/1 Wellcome Trust 208385/Z/17/Z BBSRC (Biotechnology & Biological Sciences Research Council) BB/M012573/1 BBSRC (Biotechnology & Biological Sciences Research Council) BB/R000271/1
Depositing User:	Symplectic Publications
Date Deposited:	19 Mar 2020 12:06
Last Modified:	19 Dec 2024 15:02
Status:	Published
Publisher:	Nature Research
Identification Number:	10.1038/s41467-020-15702-1
Related URLs:	Dataset
Open Archives Initiative ID (OAI ID):	oai:eprints.whiterose.ac.uk:158540