Small-angle neutron scattering from the reconstituted TF1 of H(+)-ATPase from thermophilic bacterium PS3 with deuterated subunits.
Subunits alpha, beta and gamma of adenosine triphosphatase (H(+)-ATPase) from the thermophilic bacterium PS3 (TF1) have been over-expressed in Escherichia coli. alpha and beta subunits deuterated to the level of 90% were obtained by culturing E. coli in 2H2O medium. Both the subunits and the reconstituted alpha beta gamma complex, TF1, which contain the deuterated components in various combinations, were studied in solution by small-angle neutron scattering. The individual shapes of the subunits and their organization in the alpha beta gamma-TF1 complex were examined using the techniques of selective deuteration and contrast variation. The alpha and beta subunits are well approximated as ellipsoids of revolution having minor semi-axes of 20.4(+/- 0.4) and 20.0(+/- 0.2) A, and major semi-axes of 53.0(+/- 1.4) and 55.8(+/- 0.9) A, respectively. In the TF1 complex, three beta subunits are aligned to form an equilateral triangle, with their major axes tilted by 35 degrees with respect to the 3-fold axis of the complex. The beta-beta distance is about 53 A. Three alpha subunits are similarly arranged, positioned between the beta subunits, and with their direction of tilt opposite to that of the beta subunits. The centers of the alpha and beta subunits lie in the same plane, forming a hexagon. Adjacent subunits overlap in this model, suggesting that they are not simple ellipsoids of revolution.
Ito
Y
Y
Institute for Solid State Physics, University of Tokyo, Japan.
Harada
M
M
Ohta
S
S
Kagawa
Y
Y
Aono
O
O
Schefer
J
J
Schoenborn
B P
BP
D001419:Bacteria / Q000201:enzymology*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003903:Deuterium; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D009502:Neutrons; D006180:Proton-Translocating ATPases / Q000378:metabolism; D012542:Scattering, Radiation
D003903:Deuterium; D006180:Proton-Translocating ATPases