Surfactosomes: a novel approach to the reconstitution and 2-D crystallisation of membrane proteins.
The formation of vesicle-like structures (termed surfactosomes) and lamellar sheets from solutions containing ammonium perfluoroocanoate (APFO) is illustrated using conventional and cryo-transmission electron microscopy. It is shown how this detergent can be used for the solubilisation, reconstitution and 2-D crystallisation of membrane proteins as demonstrated for the major protein of the membrane sector of the V-type H+-ATPase (16-kDa protein). Electron microscopical analysis of 2-D crystals of the 16-kDa protein (a=b=13.0+/-0.2nm with gamma = 90 degrees and p4 projection symmetry) revealed a unit cell comprising four dimeric complexes of the 16-kDa protein the significance of which is discussed.
Reviakine
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I
Department of Biology, University of Leeds, UK.
Stoylova
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S
Holzenburg
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A
D000818:Animals; D002210:Caprylates; D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D005466:Fluorocarbons; D008081:Liposomes / Q000737:chemistry*; D008099:Liver / Q000201:enzymology; D008565:Membrane Proteins / Q000737:chemistry*; D008854:Microscopy, Electron; D008121:Nephropidae; D010179:Pancreas / Q000201:enzymology; D006180:Proton-Translocating ATPases / Q000737:chemistry; D012995:Solubility; D013501:Surface-Active Agents
D002210:Caprylates; D005466:Fluorocarbons; D008081:Liposomes; D008565:Membrane Proteins; D013501:Surface-Active Agents; C023036:perfluorooctanoic acid; D006180:Proton-Translocating ATPases