Formation in vivo, purification and crystallization of a complex of the gamma and epsilon subunits of the F0F1-ATPase of Escherichia coli.
A complex comprising the epsilon subunit of Escherichia coli F1-ATPase (ECF1-ATPase) and a glutathione-S-transferase gamma subunit (of ECF1-ATPase) fusion protein was formed in vivo and purified from cell extracts by binding to glutathione-agarose beads. The glutathione-S-transferase was released from the complex by digestion with thrombin and the gamma/epsilon complex purified by cation-exchange chromatography. Crystals of the complex were grown by vapour diffusion using PEG8000 as precipitant. The crystals are orthorhombic, space-group P2(1)2(1)2 with a = 161.9 A, b = 44.1 A and c = 63.4 A. The volume of the asymmetric unit is consistent with the presence of a complex of one gamma subunit and one epsilon subunit.
Cox
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GB
Department of Biochemistry and Molecular Biology, John Curtin School of Medical Research, Australian National University, Canberra City, A.C.T.
Cromer
B A
BA
Guss
J M
JM
Harvey
I
I
Jeffrey
P D
PD
Solomon
R G
RG
Webb
D C
DC
D002852:Chromatography, Ion Exchange; D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D005982:Glutathione Transferase / Q000235:genetics; D010957:Plasmids; D006180:Proton-Translocating ATPases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics
D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase; D006180:Proton-Translocating ATPases