Selection of membrane protein targets for crystallization using PFO-PAGE electrophoresis.
A method to rapidly assess the oligomeric composition of multimeric proteins is notably absent from reported schemes for high throughput production and crystallization of membrane proteins. In this report we have investigated the suitability of PFO-PAGE electrophoresis for this purpose and present examples where it proves highly informative in selecting conditions favouring the functional oligomeric state of the target protein. Features such as the ability to analyze several samples in parallel, including crude membrane extracts, suggest it will be highly adaptable to high throughput analysis of membrane proteins.
Cleverley
R M
RM
Manchester Interdisciplinary Biocentre, University of Manchester, Manchester, UK. Robert.cleverley@manchester.ac.uk
Saleem
M
M
Kean
J
J
Ford
R C
RC
Derrick
J P
JP
Prince
S M
SM
D018528:ATP-Binding Cassette Transporters / Q000737:chemistry / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification*; D002210:Caprylates; D027682:Cation Transport Proteins / Q000737:chemistry / Q000302:isolation & purification; D002462:Cell Membrane / Q000737:chemistry; D002850:Chromatography, Gel; D003460:Crystallization; D003902:Detergents / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005466:Fluorocarbons; D026901:Membrane Transport Proteins / Q000737:chemistry* / Q000302:isolation & purification; D020836:Protein Structure, Quaternary
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D002210:Caprylates; D027682:Cation Transport Proteins; D003902:Detergents; D005466:Fluorocarbons; D026901:Membrane Transport Proteins; C023036:perfluorooctanoic acid