Conformational Dynamics of AcrA Govern Multidrug Efflux Pump Assembly

Hazel, Anthony J.; Abdali, Narges; Leus, Inga V.; Parks, Jerry M.; Smith, Jeremy C.; Zgurskaya, Helen I.; Gumbart, James C.

doi:10.1021/acsinfecdis.9b00273

Title: Conformational Dynamics of AcrA Govern Multidrug Efflux Pump Assembly

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Abstract

Multidrug efflux pumps of pathogenic, Gram-negative bacteria comprise an innate resistance mechanism and are key contributors to the emerging global pandemic of antibiotic resistance. Several increasingly detailed cryo-electron microscopy maps have been resolved of an entire efflux pump complex, AcrAB–TolC, resulting in atomistic structural models. Using a recent model, we have carried out nearly 40 μs of molecular dynamics simulations to study one of the key components of the protein complex AcrA, the membrane fusion protein that connects the inner-membrane-bound AcrB to the outer-membrane-bound TolC. Here, we determined a three-dimensional potential of mean force (PMF) for AcrA, which displays two main conformational basins representing assembly competent and incompetent states. Corresponding experiments show that stabilizing mutations at an interdomain interface shift the dynamic equilibrium between these states to the incompetent one, disrupting pump assembly and function and resensitizing bacteria to existing antibiotics. The modulation of AcrA dynamics through pharmacological intervention therefore presents a promising route for the development of new antibiotics.

Authors:

Hazel, Anthony J. ^[1]; Abdali, Narges ^[2]; Leus, Inga V. ^[2];

^[3];

^[4];

^[2];

^[1]

Georgia Inst. of Technology, Atlanta, GA (United States)
Univ. of Oklahoma, Norman, OK (United States)
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Univ. of Tennessee, Knoxville, TN (United States)

Publication Date:: Fri Sep 13 00:00:00 EDT 2019

Research Org.:: Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

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

OSTI Identifier:: 1607026

Grant/Contract Number:: AC05-00OR22725; R01-AI052293; R01-GM123169; OCI-1053575

Resource Type:: Accepted Manuscript

Journal Name:: ACS Infectious Diseases

Additional Journal Information:: Journal Volume: 5; Journal Issue: 11; Journal ID: ISSN 2373-8227

Publisher:: American Chemical Society (ACS)

Country of Publication:: United States

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES; antibiotic resistance; efflux pump; Gram-negative bacteria; molecular dynamics simulations; free-energy calculations

Citation Formats


                    Hazel, Anthony J., Abdali, Narges, Leus, Inga V., Parks, Jerry M., Smith, Jeremy C., Zgurskaya, Helen I., and Gumbart, James C. Conformational Dynamics of AcrA Govern Multidrug Efflux Pump Assembly.  United States: N. p., 2019. 
Web.  doi:10.1021/acsinfecdis.9b00273.

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                    Hazel, Anthony J., Abdali, Narges, Leus, Inga V., Parks, Jerry M., Smith, Jeremy C., Zgurskaya, Helen I., & Gumbart, James C. Conformational Dynamics of AcrA Govern Multidrug Efflux Pump Assembly.  United States.  https://doi.org/10.1021/acsinfecdis.9b00273

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                    Hazel, Anthony J., Abdali, Narges, Leus, Inga V., Parks, Jerry M., Smith, Jeremy C., Zgurskaya, Helen I., and Gumbart, James C. Fri .  
"Conformational Dynamics of AcrA Govern Multidrug Efflux Pump Assembly".  United States.  https://doi.org/10.1021/acsinfecdis.9b00273.  https://www.osti.gov/servlets/purl/1607026.

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@article{osti_1607026,

  title        = {Conformational Dynamics of AcrA Govern Multidrug Efflux Pump Assembly},

  author       = {Hazel, Anthony J. and Abdali, Narges and Leus, Inga V. and Parks, Jerry M. and Smith, Jeremy C. and Zgurskaya, Helen I. and Gumbart, James C.},

  abstractNote = {Multidrug efflux pumps of pathogenic, Gram-negative bacteria comprise an innate resistance mechanism and are key contributors to the emerging global pandemic of antibiotic resistance. Several increasingly detailed cryo-electron microscopy maps have been resolved of an entire efflux pump complex, AcrAB–TolC, resulting in atomistic structural models. Using a recent model, we have carried out nearly 40 μs of molecular dynamics simulations to study one of the key components of the protein complex AcrA, the membrane fusion protein that connects the inner-membrane-bound AcrB to the outer-membrane-bound TolC. Here, we determined a three-dimensional potential of mean force (PMF) for AcrA, which displays two main conformational basins representing assembly competent and incompetent states. Corresponding experiments show that stabilizing mutations at an interdomain interface shift the dynamic equilibrium between these states to the incompetent one, disrupting pump assembly and function and resensitizing bacteria to existing antibiotics. The modulation of AcrA dynamics through pharmacological intervention therefore presents a promising route for the development of new antibiotics.},

  doi          = {10.1021/acsinfecdis.9b00273},

  journal      = {ACS Infectious Diseases},

  number       = 11,

  volume       = 5,

  place        = {United States},

  year         = {Fri Sep 13 00:00:00 EDT 2019},

  month        = {Fri Sep 13 00:00:00 EDT 2019}

}

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