Using rational screening and electron microscopy to optimize the crystallization of succinate:ubiquinone oxidoreductase from Escherichia coli.
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.
Horsefield
Rob
R
Department of Biological Sciences, Imperial College, London SW7 2AZ, England.
Yankovskaya
Victoria
V
Törnroth
Susanna
S
Luna-Chavez
César
C
Stambouli
Elizabeth
E
Barber
James
J
Byrne
Bernadette
B
Cecchini
Gary
G
Iwata
So
S
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
D004396:Coloring Agents; D009097:Multienzyme Complexes; D010088:Oxidoreductases; D042963:Electron Transport Complex II; D013385:Succinate Dehydrogenase