title: Microbial drug efflux proteins of the major facilitator superfamily.
abstract: Drug efflux proteins are widespread amongst microorganisms, including pathogens. They can contribute to both natural insensitivity to antibiotics and to emerging antibiotic resistance and so are potential targets for the development of new antibacterial drugs. The design of such drugs would be greatly facilitated by knowledge of the structures of these transport proteins, which are poorly understood, because of the difficulties of obtaining crystals of quality. We describe a structural genomics approach for the amplified expression, purification and characterisation of prokaryotic drug efflux proteins of the 'Major Facilitator Superfamily' (MFS) of transport proteins from Helicobacter pylori, Staphylococcus aureus, Escherichia coli, Enterococcus faecalis, Bacillus subtilis, Brucella melitensis, Campylobacter jejuni, Neisseria meningitides and Streptomyces coelicolor. The H. pylori putative drug resistance protein, HP1092, and the S. aureus QacA proteins are used as detailed examples. This strategy is an important step towards reproducible production of transport proteins for the screening of drug binding and for optimisation of crystallisation conditions to enable subsequent structure determination.
authors.affiliation: Astbury Centre for Structural Molecular Biology, School of Biochemistry and Microbiology, University of Leeds, Leeds LS2 9JT, UK.
authors.forename: Massoud
authors.initials: M
authors.lastname: Saidijam
authors.forename: Giulia
authors.initials: G
authors.lastname: Benedetti
authors.forename: Qinghu
authors.initials: Q
authors.lastname: Ren
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Xu
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Hoyle
authors.forename: Sarah L
authors.initials: SL
authors.lastname: Palmer
authors.forename: Alison
authors.initials: A
authors.lastname: Ward
authors.forename: Kim E
authors.initials: KE
authors.lastname: Bettaney
authors.forename: Gerda
authors.initials: G
authors.lastname: Szakonyi
authors.forename: Johan
authors.initials: J
authors.lastname: Meuller
authors.forename: Scott
authors.initials: S
authors.lastname: Morrison
authors.forename: Martin K
authors.initials: MK
authors.lastname: Pos
authors.forename: Patrick
authors.initials: P
authors.lastname: Butaye
authors.forename: Karl
authors.initials: K
authors.lastname: Walravens
authors.forename: Kate
authors.initials: K
authors.lastname: Langton
authors.forename: Richard B
authors.initials: RB
authors.lastname: Herbert
authors.forename: Ronald A
authors.initials: RA
authors.lastname: Skurray
authors.forename: Ian T
authors.initials: IT
authors.lastname: Paulsen
authors.forename: John
authors.initials: J
authors.lastname: O'reilly
authors.forename: Nicholas G
authors.initials: NG
authors.lastname: Rutherford
authors.forename: Melissa H
authors.initials: MH
authors.lastname: Brown
authors.forename: Roslyn M
authors.initials: RM
authors.lastname: Bill
authors.forename: Peter J F
authors.initials: PJ
authors.lastname: Henderson
mesh_terms: D000595:Amino Acid Sequence; D001419:Bacteria / Q000235:genetics / Q000378:metabolism*; D024881:Drug Resistance, Bacterial; D015964:Gene Expression Regulation, Bacterial; D026901:Membrane Transport Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017510:Protein Folding
chemical_list: D026901:Membrane Transport Proteins