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BpeB, a major resistance‐nodulation‐cell division transporter from Burkholderia cenocepacia : construct design, crystallization and preliminary structural analysis
Author(s) -
Horikawa Tomonari,
Hung Li-Wei,
Kim Heung-Bok,
Shaya David,
Kim Chang-Yub,
Terwilliger Thomas C.,
Yamashita Eiki,
Aoki Maho,
Okada Ui,
Murakami Satoshi
Publication year - 2018
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/s2053230x18013547
Subject(s) - burkholderia cenocepacia , efflux , biology , microbiology and biotechnology , transporter , burkholderia , biochemistry , bacteria , genetics , gene
Burkholderia cenocepacia is an opportunistic pathogen that infects cystic fibrosis patients, causing pneumonia and septicemia. B. cenocepacia has intrinsic antibiotic resistance against monobactams, aminoglycosides, chloramphenicol and fluoroquinolones that is contributed by a homologue of BpeB, which is a member of the resistance‐nodulation‐cell division (RND)‐type multidrug‐efflux transporters. Here, the cloning, overexpression, purification, construct design for crystallization and preliminary X‐ray diffraction analysis of this BpeB homologue from B. cenocepacia are reported. Two truncation variants were designed to remove possible disordered regions based on comparative sequence and structural analysis to salvage the wild‐type protein, which failed to crystallize. The 17‐residue carboxyl‐terminal truncation yielded crystals that diffracted to 3.6 Å resolution. The efflux function measured using minimal inhibitory concentration assays indicated that the truncation decreased, but did not eliminate, the efflux activity of the transporter.