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Measuring Small Molecule Binding to Escherichia coli AcrB by Surface Plasmon Resonance

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Targeting Enzymes for Pharmaceutical Development

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2089))

Abstract

Antimicrobial resistance (AMR) is rapidly becoming one of the great healthcare challenges. A common mechanism employed by pathogenic bacteria to avoid the action of certain antibiotics is to overexpress efflux pumps that can extrude these drugs from the cell rendering them ineffective. Small molecule inhibitors that target bacterial efflux pumps provide a route toward reversing AMR. Here, we describe the application of surface plasmon resonance (SPR) technology to characterize protein:small molecule interactions between the inner membrane protein AcrB subunit of the Escherichia coli AcrA-AcrB-TolC efflux pump and its substrates and novel inhibitors. The SPR assay provides quantitative data about the kinetics of binding that can help guide the development of new chemotherapies to combat AMR.

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Acknowledgments

This research was supported financially by the National Natural Science Foundation of China (81673284), National Health and Medical Research Council of Australia (GN1147538), China–Australia Centre for Health Sciences Research and Australian Postgraduate Award (RM). All experiments were performed on a BiaCore 200 which from part of the Biophysical Characterisation Suite at The University of South Australia.

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Authors and Affiliations

  1. School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, SA, Australia

    Steven W. Polyak, Rumana Mowla & Henrietta Venter

Authors
  1. Steven W. Polyak
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  2. Rumana Mowla
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  3. Henrietta Venter
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Corresponding author

Correspondence to Henrietta Venter .

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Editors and Affiliations

  1. Department of Biotechnology, Agricultural University of Athens, Athens, Greece

    Nikolaos E. Labrou

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Polyak, S.W., Mowla, R., Venter, H. (2020). Measuring Small Molecule Binding to Escherichia coli AcrB by Surface Plasmon Resonance. In: Labrou, N. (eds) Targeting Enzymes for Pharmaceutical Development. Methods in Molecular Biology, vol 2089. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0163-1_7

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  • DOI: https://doi.org/10.1007/978-1-0716-0163-1_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0162-4

  • Online ISBN: 978-1-0716-0163-1

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