Crystallographic Analysis of MATE-Type Multidrug Exporter with Its Inhibitors
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 wordsMultidrug resistance Multidrug exporter Transporter Membrane proteins X-ray crystallography Structural analysis Macrocyclic peptide Inhibitors RaPID system
- 6.McAleese F, Petersen P, Ruzin A et al (2005) A novel MATE family efflux pump contributes to the reduced susceptibility of laboratory-derived Staphylococcus aureus mutants to tigecycline. Antimicrob Agents Chemother 49:1865–1871. https://doi.org/10.1128/AAC.49.5.1865-1871.2005 CrossRefPubMedPubMedCentralGoogle Scholar
- 25.Eswar N, Webb B, Marti-Renom MA et al (2007) Comparative protein structure modeling using MODELLER. Curr Protoc Protein Sci. https://doi.org/10.1002/0471140864.ps0209s50