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

Calabrese, Antonio N. and Schiffrin, Bob and Watson, Matthew and Walko, Martin and Humes, Julia R. and Karamanos, Theodoros K. and Horne, Jim E. and White, Paul and Kalli, Antreas C. and Tuma, Roman and Ashcroft, Alison E. and Brockwell, David J. and Radford, Sheena E. (2020) Inter-domain dynamics in the chaperone SurA and multi-site binding to its outer membrane protein clients - dataset. University of Leeds. [Dataset] https://doi.org/10.5518/701

Dataset description

SurA is the conserved major chaperone of outer membrane protein (OMP) biogenesis in the periplasm of Gram-negative bacteria, and plays a key role in cell envelope homeostasis and virulence. In E. coli, SurA comprises three domains: a core domain and two peptidyl prolyl isomerase (PPIase) domains (P1 and P2). The functional roles of these domains in chaperone activity, including their inter-subunit dynamics and interactions with substrates remain unclear. Here we have used chemical cross-linking, HDX-MS, oxidative labelling, single-molecule FRET and molecular dynamics simulations, to investigate the conformational dynamics of SurA in its apo state and to map the binding site(s) for its clients. We demonstrate that SurA predominantly samples conformations in solution in which the P2 domain is closer to the core and P1 domains than suggested by the extended conformation in its crystal structure. Using cross-linking experiments we show that SurA has multiple, yet specific, binding sites for OmpX located primarily in the core domain, and that residues spanning the whole OmpX sequence bind in these regions. HDX experiments additionally reveal that binding of OmpX, and the larger client OmpF, lead to inter-domain conformational changes between the core and P1 domains. Combining these data we generate a model for SurA OMP recognition in which unfolded OMP substrates bind in a cradle formed between the SurA domains, in a manner reminiscent of other ATP-independent chaperones. Structural flexibility between domains is likely a key feature of the SurA functional cycle, assisting in OMP recognition, binding and release of its broad range of OMP substrates for folding into the outer membrane.

Additional information: Equipment used: 105615/Z/14/Z 208385/Z/17/Z BB/M012573/1
Keywords: Biological science, Protein structure and dynamics
Subjects: C000 - Biological sciences
Related resources:
LocationType
https://doi.org/10.1101/2019.12.19.882696Publication
http://eprints.whiterose.ac.uk/158540/Publication
License: Creative Commons Attribution 4.0 International (CC BY 4.0)
Date deposited: 09 Mar 2020 21:07
URI: https://archive.researchdata.leeds.ac.uk/id/eprint/650

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