title: Using rational screening and electron microscopy to optimize the crystallization of succinate:ubiquinone oxidoreductase from Escherichia coli.
abstract: The membrane-bound respiratory complex II, succinate:ubiquinone oxidoreductase (SQR) from Escherichia coli, has been anaerobically expressed, then purified and crystallized. The initial crystals obtained were small and diffracted poorly. In order to facilitate structure determination, rational screening and sample-quality analysis using electron microscopy was implemented. The crystals of SQR from E. coli belong to the trigonal space group R32, with unit-cell parameters a = b = 138.7, c = 521.9 A, and diffract to 2.6 A resolution. The optimization strategy used for obtaining well diffracting SQR crystals is applicable to a wide range of membrane proteins.
authors.affiliation: Department of Biological Sciences, Imperial College, London SW7 2AZ, England.
authors.forename: Rob
authors.initials: R
authors.lastname: Horsefield
authors.forename: Victoria
authors.initials: V
authors.lastname: Yankovskaya
authors.forename: Susanna
authors.initials: S
authors.lastname: Törnroth
authors.forename: César
authors.initials: C
authors.lastname: Luna-Chavez
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Stambouli
authors.forename: James
authors.initials: J
authors.lastname: Barber
authors.forename: Bernadette
authors.initials: B
authors.lastname: Byrne
authors.forename: Gary
authors.initials: G
authors.lastname: Cecchini
authors.forename: So
authors.initials: S
authors.lastname: Iwata
mesh_terms: D016880:Anisotropy; D004396:Coloring Agents; D003460:Crystallization; D018360:Crystallography, X-Ray; D042963:Electron Transport Complex II; D004926:Escherichia coli / Q000201:enzymology*; D008854:Microscopy, Electron; D009097:Multienzyme Complexes / Q000737:chemistry* / Q000302:isolation & purification; D010088:Oxidoreductases / Q000737:chemistry* / Q000302:isolation & purification; D013385:Succinate Dehydrogenase / Q000737:chemistry* / Q000302:isolation & purification; D014461:Ultracentrifugation
chemical_list: D004396:Coloring Agents; D009097:Multienzyme Complexes; D010088:Oxidoreductases; D042963:Electron Transport Complex II; D013385:Succinate Dehydrogenase