BpeB, a major resistance-nodulation-cell division transporter from Burkholderia cenocepacia: construct design, crystallization and preliminary structural analysis.
Tomonari HorikawaLi-Wei HungHeung Bok KimDavid ShayaChang Yub KimThomas C TerwilligerEiki YamashitaMaho AokiUi OkadaSatoshi MurakamiPublished in: Acta crystallographica. Section F, Structural biology communications (2018)
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.
Keyphrases
- cystic fibrosis
- end stage renal disease
- wild type
- single cell
- cell therapy
- chronic kidney disease
- ejection fraction
- newly diagnosed
- amino acid
- prognostic factors
- cell proliferation
- high throughput
- peritoneal dialysis
- drug resistant
- high resolution
- copy number
- stem cells
- mesenchymal stem cells
- candida albicans
- mass spectrometry
- computed tomography
- air pollution
- intensive care unit
- patient reported outcomes
- chronic obstructive pulmonary disease
- extracorporeal membrane oxygenation
- acute respiratory distress syndrome
- dual energy