title: Collection and characterisation of bacterial membrane proteins.
abstract: A general strategy for the amplified expression in Escherichia coli of membrane transport and receptor proteins from other bacteria is described. As an illustration we report the cloning of the putative alpha-ketoglutarate membrane transport gene from the genome of Helicobacter pylori, overexpression of the protein tagged with RGS(His)6 at the C-terminus, and its purification in mg quantities. The retention of structural and functional integrity was verified by circular dichroism spectroscopy and reconstitution of transport activity. This strategy for overexpression and purification is extended to additional membrane proteins from H. pylori and from other bacteria.
authors: Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
authors: Massoud
authors: M
authors: Saidijam
authors: Georgios
authors: G
authors: Psakis
authors: Joanne L
authors: JL
authors: Clough
authors: Johan
authors: J
authors: Meuller
authors: Shun'ichi
authors: S
authors: Suzuki
authors: Christopher J
authors: CJ
authors: Hoyle
authors: Sarah L
authors: SL
authors: Palmer
authors: Scott M
authors: SM
authors: Morrison
authors: Martin K
authors: MK
authors: Pos
authors: Richard C
authors: RC
authors: Essenberg
authors: Martin C J
authors: MC
authors: Maiden
authors: Atif
authors: A
authors: Abu-bakr
authors: Simon G
authors: SG
authors: Baumberg
authors: Alex A
authors: AA
authors: Neyfakh
authors: Jeffrey K
authors: JK
authors: Griffith
authors: Michael J
authors: MJ
authors: Stark
authors: Alison
authors: A
authors: Ward
authors: John
authors: J
authors: O'Reilly
authors: Nicholas G
authors: NG
authors: Rutherford
authors: Mary K
authors: MK
authors: Phillips-Jones
authors: Peter J F
authors: PJ
authors: Henderson
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000302:isolation & purification*; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics / Q000302:isolation & purification; D002942:Circular Dichroism; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004274:DNA, Recombinant / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D005822:Genetic Vectors; D016480:Helicobacter pylori / Q000235:genetics; D008565:Membrane Proteins / Q000235:genetics* / Q000302:isolation & purification*; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D012995:Solubility
chemical_list: D001426:Bacterial Proteins; D002352:Carrier Proteins; D004269:DNA, Bacterial; D004274:DNA, Recombinant; D008565:Membrane Proteins; D011994:Recombinant Proteins