title: Active membrane transport and receptor proteins from bacteria.
abstract: A general strategy for the expression of bacterial membrane transport and receptor genes in Escherichia coli is described. Expression is amplified so that the encoded proteins comprise 5-35% of E. coli inner membrane protein. Depending upon their topology, proteins are produced with RGSH6 or a Strep tag at the C-terminus. These enable purification in mg quantities for crystallization and NMR studies. Examples of one nutrient uptake and one multidrug extrusion protein from Helicobacter pylori are described. This strategy is successful for membrane proteins from H. pylori, E. coli, Enterococcus faecalis, Bacillus subtilis, Staphylococcus aureus, Microbacterium liquefaciens, Brucella abortus, Brucella melitensis, Campylobacter jejuni, Neisseria meningitides, Streptomyces coelicolor and Rhodobacter sphaeroides.
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, U.K.
authors.forename: M
authors.initials: M
authors.lastname: Saidijam
authors.forename: K E
authors.initials: KE
authors.lastname: Bettaney
authors.forename: G
authors.initials: G
authors.lastname: Szakonyi
authors.forename: G
authors.initials: G
authors.lastname: Psakis
authors.forename: K
authors.initials: K
authors.lastname: Shibayama
authors.forename: S
authors.initials: S
authors.lastname: Suzuki
authors.forename: J L
authors.initials: JL
authors.lastname: Clough
authors.forename: V
authors.initials: V
authors.lastname: Blessie
authors.forename: A
authors.initials: A
authors.lastname: Abu-Bakr
authors.forename: S
authors.initials: S
authors.lastname: Baumberg
authors.forename: J
authors.initials: J
authors.lastname: Meuller
authors.forename: C K
authors.initials: CK
authors.lastname: Hoyle
authors.forename: S L
authors.initials: SL
authors.lastname: Palmer
authors.forename: P
authors.initials: P
authors.lastname: Butaye
authors.forename: K
authors.initials: K
authors.lastname: Walravens
authors.forename: S G
authors.initials: SG
authors.lastname: Patching
authors.forename: J
authors.initials: J
authors.lastname: O'reilly
authors.forename: N G
authors.initials: NG
authors.lastname: Rutherford
authors.forename: R M
authors.initials: RM
authors.lastname: Bill
authors.forename: D I
authors.initials: DI
authors.lastname: Roper
authors.forename: M K
authors.initials: MK
authors.lastname: Phillips-Jones
authors.forename: P J F
authors.initials: PJ
authors.lastname: Henderson
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001693:Biological Transport, Active; D002942:Circular Dichroism; D004269:DNA, Bacterial / Q000235:genetics; D016480:Helicobacter pylori / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008954:Models, Biological; D010957:Plasmids; D015183:Restriction Mapping; D027981:Symporters / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D026901:Membrane Transport Proteins; D027981:Symporters