Crystallographic Analysis of MATE-Type Multidrug Exporter with Its Inhibitors

  • Tsukasa Kusakizako
  • Yoshiki Tanaka
  • Christopher J. Hipolito
  • Hiroaki Suga
  • Osamu Nureki
Part of the Methods in Molecular Biology book series (MIMB, volume 1700)


Multidrug exporters expressed in pathogens efflux substrate drugs such as antibiotics, and thus, the development of inhibitors against them has eagerly been anticipated. Furthermore, the crystal structures of multidrug exporters with their inhibitors provide novel insights into the inhibitory mechanism and the development of more specific and effective inhibitors. We previously reported the complex structures of the Multidrug And Toxic compound Extrusion (MATE)-type multidrug exporter with the macrocyclic peptides, which inhibit the efflux of substrates by the MATE-type multidrug exporter (Tanaka et al., Nature 496:247–251, 2013). In this chapter, we describe methodologies of the screening and synthesis of macrocyclic peptides as inhibitors, as well as the purification, crystallization, and structure determination of the complexes of the MATE-type multidrug exporter with its inhibitors.

Key words

Multidrug resistance Multidrug exporter Transporter Membrane proteins X-ray crystallography Structural analysis Macrocyclic peptide Inhibitors RaPID system 


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Copyright information

© Springer Science+Business Media LLC 2018

Authors and Affiliations

  • Tsukasa Kusakizako
    • 1
  • Yoshiki Tanaka
    • 2
  • Christopher J. Hipolito
    • 3
    • 4
  • Hiroaki Suga
    • 3
  • Osamu Nureki
    • 1
  1. 1.Department of Biological Sciences, Graduate School of ScienceThe University of TokyoTokyoJapan
  2. 2.Department of Systems Biology, Graduate School of Biological SciencesNara Institute of Science and TechnologyNaraJapan
  3. 3.Department of Chemistry, Graduate School of ScienceThe University of TokyoTokyoJapan
  4. 4.Faculty of MedicineUniversity of TsukubaTsukubaJapan

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