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

Conformational restriction shapes the inhibition of a multidrug efflux adaptor protein

Journal Article · Tue Jul 18 00:00:00 EDT 2023 · Nature Communications

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

; Uddin, Muhammad R.; ; Kuo, Katie M.; Higgins, Anna J.; ; ; ; ; ; ;

Abstract

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

Grant/Contract Number:: AC05-00OR22725

OSTI ID:: 1991148

Alternate ID(s):: OSTI ID: 1991730

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

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United Kingdom

Language:: English

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