Microbial drug efflux proteins of the major facilitator superfamily.
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.
Saidijam
Massoud
M
Astbury Centre for Structural Molecular Biology, School of Biochemistry and Microbiology, University of Leeds, Leeds LS2 9JT, UK.
Benedetti
Giulia
G
Ren
Qinghu
Q
Xu
Zhiqiang
Z
Hoyle
Christopher J
CJ
Palmer
Sarah L
SL
Ward
Alison
A
Bettaney
Kim E
KE
Szakonyi
Gerda
G
Meuller
Johan
J
Morrison
Scott
S
Pos
Martin K
MK
Butaye
Patrick
P
Walravens
Karl
K
Langton
Kate
K
Herbert
Richard B
RB
Skurray
Ronald A
RA
Paulsen
Ian T
IT
O'reilly
John
J
Rutherford
Nicholas G
NG
Brown
Melissa H
MH
Bill
Roslyn M
RM
Henderson
Peter J F
PJ
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
D026901:Membrane Transport Proteins