Conformational restriction shapes the inhibition of a multidrug efflux adaptor protein

Russell Lewis, Benjamin; Uddin, Muhammad R.; Moniruzzaman, Mohammad; Kuo, Katie M.; Higgins, Anna J.; Shah, Laila M. N.; Sobott, Frank; Parks, Jerry M.; Hammerschmid, Dietmar; Gumbart, James C.; Zgurskaya, Helen I.; Reading, Eamonn

doi:10.1038/s41467-023-39615-x

Title: Conformational restriction shapes the inhibition of a multidrug efflux adaptor protein

Journal Article · 18 July 2023 · Nature Communications

DOI: https://doi.org/10.1038/s41467-023-39615-x · OSTI ID:1991148

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King's College, London (United Kingdom)
Univ. of Oklahoma, Norman, OK (United States)
Georgia Institute of Technology, Atlanta, GA (United States)
University of Leeds (United Kingdom)
Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)

Membrane efflux pumps play a major role in bacterial multidrug resistance. The tripartite multidrug efflux pump system from Escherichia coli, AcrAB-TolC, is a target for inhibition to lessen resistance development and restore antibiotic efficacy, with homologs in other ESKAPE pathogens. Here, we rationalize a mechanism of inhibition against the periplasmic adaptor protein, AcrA, using a combination of hydrogen/deuterium exchange mass spectrometry, cellular efflux assays, and molecular dynamics simulations. We define the structural dynamics of AcrA and find that an inhibitor can inflict long-range stabilisation across all four of its domains, whereas an interacting efflux substrate has minimal effect. Our results support a model where an inhibitor forms a molecular wedge within a cleft between the lipoyl and αβ barrel domains of AcrA, diminishing its conformational transmission of drug-evoked signals from AcrB to TolC. This work provides molecular insights into multidrug adaptor protein function which could be valuable for developing antimicrobial therapeutics.

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Research Organization:: Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)

Sponsoring Organization:: USDOE; National Institutes of Health (NIH); National Science Foundation (NSF)

Grant/Contract Number:: AC05-00OR22725; R01-AI052293; ACI-1548562; 1828187

OSTI ID:: 1991148

Alternate ID(s):: OSTI ID: 1991730

Journal Information:: Nature Communications, Vol. 14, Issue 1; ISSN 2041-1723

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

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