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LCP crystallization and X‐ray diffraction analysis of VcmN, a MATE transporter from Vibrio cholerae
Author(s) -
Kusakizako Tsukasa,
Tanaka Yoshiki,
Hipolito Christopher J.,
Kuroda Teruo,
Ishitani Ryuichiro,
Suga Hiroaki,
Nureki Osamu
Publication year - 2016
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x16008931
Subject(s) - vibrio cholerae , crystallization , microbiology and biotechnology , transporter , biology , chemistry , bacteria , genetics , gene , organic chemistry
Multidrug and toxic compound extrusion (MATE) transporters, one of the multidrug exporter families, efflux xenobiotics towards the extracellular side of the membrane. Since MATE transporters expressed in bacterial pathogens contribute to multidrug resistance, they are important therapeutic targets. Here, a MATE‐transporter homologue from Vibrio cholerae , VcmN, was overexpressed in Escherichia coli , purified and crystallized in lipidic cubic phase (LCP). X‐ray diffraction data were collected to 2.5 Å resolution from a single crystal obtained in a sandwich plate. The crystal belonged to space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 52.3, b = 93.7, c = 100.2 Å. As a result of further LCP crystallization trials, crystals of larger size were obtained using sitting‐drop plates. X‐ray diffraction data were collected to 2.2 Å resolution from a single crystal obtained in a sitting‐drop plate. The crystal belonged to space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 61.9, b = 91.8, c = 100.9 Å. The present work provides valuable insights into the atomic resolution structure determination of membrane transporters.