The expression, purification and crystallization of the epsilon subunit of the F1 portion of the ATPase of Escherichia coli.
The epsilon subunit of the F0F1-ATPase from Escherichia coli has been expressed in E. coli as a fusion protein with glutathione S-transferase from the parasitic helminth Schistosoma japonicum. The epsilon subunit released by thrombin treatment of the purified fusion protein carried two amino acid changes, A1G and M2S, and was obtained in a yield of about five milligrams per litre of cultured cells. The two amino acid changes were shown not to affect function. The protein has been crystallized in a form suitable for X-ray diffraction structure analysis. The crystals are hexagonal, space group P6(1)22 (or P6(5)22), with a = b = 94.9 A, c = 57.1 A and gamma = 120 degrees. The diffraction from small crystals extends to at least 2.9 A resolution.
Codd
R
R
Department of Inorganic Chemistry, University of Sydney, N.S.W., Australia.
Cox
G B
GB
Guss
J M
JM
Solomon
R G
RG
Webb
D
D
D003460:Crystallization; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D005982:Glutathione Transferase / Q000235:genetics; D006180:Proton-Translocating ATPases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014961:X-Ray Diffraction
D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase; D006180:Proton-Translocating ATPases