Purification, crystallization, and preliminary X-ray crystallographic analysis of thermus thermophilus V(1)-ATPase B subunit.
The gene of V(1)-ATPase B subunit from the thermophilic eubacterium Thermus thermophilus has been cloned and the protein overproduced in Escherichia coli. The purified protein, with a molecular weight of 53.2 kDa, was crystallized from 10% (w/v) polyethylene glycol 1000, 120 mM magnesium chloride, and 100 mM Na-tricine, pH 8.0, by the vapor diffusion method. The crystals diffracted X-rays beyond 3.5 A on a synchrotron radiation source. The crystals belong to the monoclinic space group C2, with unit cell dimensions of a = 153.1 A, b = 129.6 A, c = 92.7 A, and beta = 100.3 degrees. Assuming that three or four molecules are contained in an asymmetric unit, the V(M) value is calculated as 2.8 or 2.1 A (3)/Da, respectively.
Nogi
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T
Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto, 606-8502, Japan.
Fukami
T A
TA
Ishida
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M
Yoshida
M
M
Miki
K
K
D003460:Crystallization; D018360:Crystallography, X-Ray; D011487:Protein Conformation; D017494:Proton Pumps / Q000737:chemistry / Q000302:isolation & purification; D006180:Proton-Translocating ATPases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D016963:Thermus thermophilus / Q000737:chemistry* / Q000201:enzymology*; D025262:Vacuolar Proton-Translocating ATPases
D017494:Proton Pumps; D025262:Vacuolar Proton-Translocating ATPases; D006180:Proton-Translocating ATPases