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Title: Structures and transport dynamics of a Campylobacter jejuni multidrug efflux pump

Abstract

Resistance-nodulation-cell division efflux pumps are integral membrane proteins that catalyze the export of substrates across cell membranes. Within the hydrophobe-amphiphile efflux subfamily, these resistance-nodulation-cell division proteins largely form trimeric efflux pumps. The drug efflux process has been proposed to entail a synchronized motion between subunits of the trimer to advance the transport cycle, leading to the extrusion of drug molecules. Here we use X-ray crystallography and single-molecule fluorescence resonance energy transfer imaging to elucidate the structures and functional dynamics of the Campylobacter jejuni CmeB multidrug efflux pump. We find that the CmeB trimer displays a very unique conformation. A direct observation of transport dynamics in individual CmeB trimers embedded in membrane vesicles indicates that each CmeB subunit undergoes conformational transitions uncoordinated and independent of each other. On the basis of our findings and analyses, we propose a model for transport mechanism where CmeB protomers function independently within the trimer.

Authors:
; ; ; ; ; ; ; ; ; ; ; ;  [1]
  1. Cornell
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
National Institutes of Health (NIH)
OSTI Identifier:
1377903
Resource Type:
Journal Article
Journal Name:
Nature Communications
Additional Journal Information:
Journal Volume: 8; Journal Issue: 1; Journal ID: ISSN 2041-1723
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES

Citation Formats

Su, Chih-Chia, Yin, Linxiang, Kumar, Nitin, Dai, Lei, Radhakrishnan, Abhijith, Bolla, Jani Reddy, Lei, Hsiang-Ting, Chou, Tsung-Han, Delmar, Jared A., Rajashankar, Kanagalaghatta R., Zhang, Qijing, Shin, Yeon-Kyun, Yu, Edward W., and Iowa State). Structures and transport dynamics of a Campylobacter jejuni multidrug efflux pump. United States: N. p., 2017. Web. doi:10.1038/s41467-017-00217-z.
Su, Chih-Chia, Yin, Linxiang, Kumar, Nitin, Dai, Lei, Radhakrishnan, Abhijith, Bolla, Jani Reddy, Lei, Hsiang-Ting, Chou, Tsung-Han, Delmar, Jared A., Rajashankar, Kanagalaghatta R., Zhang, Qijing, Shin, Yeon-Kyun, Yu, Edward W., & Iowa State). Structures and transport dynamics of a Campylobacter jejuni multidrug efflux pump. United States. doi:10.1038/s41467-017-00217-z.
Su, Chih-Chia, Yin, Linxiang, Kumar, Nitin, Dai, Lei, Radhakrishnan, Abhijith, Bolla, Jani Reddy, Lei, Hsiang-Ting, Chou, Tsung-Han, Delmar, Jared A., Rajashankar, Kanagalaghatta R., Zhang, Qijing, Shin, Yeon-Kyun, Yu, Edward W., and Iowa State). Tue . "Structures and transport dynamics of a Campylobacter jejuni multidrug efflux pump". United States. doi:10.1038/s41467-017-00217-z.
@article{osti_1377903,
title = {Structures and transport dynamics of a Campylobacter jejuni multidrug efflux pump},
author = {Su, Chih-Chia and Yin, Linxiang and Kumar, Nitin and Dai, Lei and Radhakrishnan, Abhijith and Bolla, Jani Reddy and Lei, Hsiang-Ting and Chou, Tsung-Han and Delmar, Jared A. and Rajashankar, Kanagalaghatta R. and Zhang, Qijing and Shin, Yeon-Kyun and Yu, Edward W. and Iowa State)},
abstractNote = {Resistance-nodulation-cell division efflux pumps are integral membrane proteins that catalyze the export of substrates across cell membranes. Within the hydrophobe-amphiphile efflux subfamily, these resistance-nodulation-cell division proteins largely form trimeric efflux pumps. The drug efflux process has been proposed to entail a synchronized motion between subunits of the trimer to advance the transport cycle, leading to the extrusion of drug molecules. Here we use X-ray crystallography and single-molecule fluorescence resonance energy transfer imaging to elucidate the structures and functional dynamics of the Campylobacter jejuni CmeB multidrug efflux pump. We find that the CmeB trimer displays a very unique conformation. A direct observation of transport dynamics in individual CmeB trimers embedded in membrane vesicles indicates that each CmeB subunit undergoes conformational transitions uncoordinated and independent of each other. On the basis of our findings and analyses, we propose a model for transport mechanism where CmeB protomers function independently within the trimer.},
doi = {10.1038/s41467-017-00217-z},
journal = {Nature Communications},
issn = {2041-1723},
number = 1,
volume = 8,
place = {United States},
year = {2017},
month = {8}
}

Works referenced in this record:

Analysis of Single-Molecule FRET Trajectories Using Hidden Markov Modeling
journal, September 2006


Crystal Structure of the Multidrug Exporter MexB from Pseudomonas aeruginosa
journal, May 2009

  • Sennhauser, Gaby; Bukowska, Magdalena A.; Briand, Christophe
  • Journal of Molecular Biology, Vol. 389, Issue 1
  • DOI: 10.1016/j.jmb.2009.04.001

Unsynchronised subunit motion in single trimeric sodium-coupled aspartate transporters
journal, October 2013

  • Erkens, Guus B.; Hänelt, Inga; Goudsmits, Joris M. H.
  • Nature, Vol. 502, Issue 7469
  • DOI: 10.1038/nature12538

Enzymatic Oxygen Scavenging for Photostability without pH Drop in Single-Molecule Experiments
journal, June 2012

  • Swoboda, Marko; Henig, Jörg; Cheng, Hsin-Mei
  • ACS Nano, Vol. 6, Issue 7
  • DOI: 10.1021/nn301895c

Coot model-building tools for molecular graphics
journal, November 2004

  • Emsley, Paul; Cowtan, Kevin
  • Acta Crystallographica Section D Biological Crystallography, Vol. 60, Issue 12, p. 2126-2132
  • DOI: 10.1107/S0907444904019158

The Health Burden of Campylobacter Infection and the Impact of Antimicrobial Resistance: Playing Chicken
journal, March 2007

  • Ruiz-Palacios, G. M.
  • Clinical Infectious Diseases, Vol. 44, Issue 5
  • DOI: 10.1086/509936

Crystal structures of the CusA efflux pump suggest methionine-mediated metal transport
journal, September 2010

  • Long, Feng; Su, Chih-Chia; Zimmermann, Michael T.
  • Nature, Vol. 467, Issue 7314
  • DOI: 10.1038/nature09395

A practical guide to single-molecule FRET
journal, May 2008

  • Roy, Rahul; Hohng, Sungchul; Ha, Taekjip
  • Nature Methods, Vol. 5, Issue 6
  • DOI: 10.1038/nmeth.1208

Crystallography & NMR System: A New Software Suite for Macromolecular Structure Determination
journal, September 1998

  • Brünger, A. T.; Adams, P. D.; Clore, G. M.
  • Acta Crystallographica Section D Biological Crystallography, Vol. 54, Issue 5
  • DOI: 10.1107/S0907444998003254

Crystal structures of a multidrug transporter reveal a functionally rotating mechanism
journal, August 2006

  • Murakami, Satoshi; Nakashima, Ryosuke; Yamashita, Eiki
  • Nature, Vol. 443, Issue 7108
  • DOI: 10.1038/nature05076

Charged Amino Acids (R83, E567, D617, E625, R669, and K678) of CusA Are Required for Metal Ion Transport in the Cus Efflux System
journal, September 2012


PHENIX : building new software for automated crystallographic structure determination
journal, October 2002

  • Adams, Paul D.; Grosse-Kunstleve, Ralf W.; Hung, Li-Wei
  • Acta Crystallographica Section D Biological Crystallography, Vol. 58, Issue 11
  • DOI: 10.1107/S0907444902016657

Crystal structure of bacterial multidrug efflux transporter AcrB
journal, October 2002

  • Murakami, Satoshi; Nakashima, Ryosuke; Yamashita, Eiki
  • Nature, Vol. 419, Issue 6907
  • DOI: 10.1038/nature01050

Alternating-Laser Excitation of Single Molecules
journal, July 2005

  • Kapanidis, Achillefs N.; Laurence, Ted A.; Lee, Nam Ki
  • Accounts of Chemical Research, Vol. 38, Issue 7
  • DOI: 10.1021/ar0401348

Crystal Structure of the Neisseria gonorrhoeae MtrD Inner Membrane Multidrug Efflux Pump
journal, June 2014


SLiCE: a novel bacterial cell extract-based DNA cloning method
journal, January 2012

  • Zhang, Yongwei; Werling, Uwe; Edelmann, Winfried
  • Nucleic Acids Research, Vol. 40, Issue 8
  • DOI: 10.1093/nar/gkr1288

Structural basis for the inhibition of bacterial multidrug exporters
journal, June 2013

  • Nakashima, Ryosuke; Sakurai, Keisuke; Yamasaki, Seiji
  • Nature, Vol. 500, Issue 7460, p. 102-106
  • DOI: 10.1038/nature12300

Mitigating Unwanted Photophysical Processes for Improved Single-Molecule Fluorescence Imaging
journal, March 2009


Coupling of remote alternating-access transport mechanisms for protons and substrates in the multidrug efflux pump AcrB
journal, September 2014


Crystal structure of the CusBA heavy-metal efflux complex of Escherichia coli
journal, February 2011

  • Su, Chih-Chia; Long, Feng; Zimmermann, Michael T.
  • Nature, Vol. 470, Issue 7335
  • DOI: 10.1038/nature09743