Mechanically driven proton conduction in single delta-free F0F1-ATPase.
In order to observe mechanically driven proton flux in F(0)F(1)-ATPase coupled with artificial driven rotation on F(1) simultaneously, a double channel observation system was established. An artificial delta-free F(0)F(1)-ATPase was constructed with alpha(3), beta(3), epsilon, gamma, and c(n) subunits as rotator and a, b(2) as stator. The chromatophore was immobilized on the glass surface through biotin-streptavidin-biotin system, and the magnetic bead was attached to the beta subunit of delta-free F(0)F(1)-ATPase. The mechanically driven proton flux was indicated by the fluorescence intensity change of fluorescein reference standard (F1300) and recorded by a cooled digital CCD camera. The mechanochemical coupling stoichiometry between F(0) and F(1) is about 4.15 +/- 0.2H(+)/rev when the magnetic field rotated at 0.33 Hz (rps).
Xiaolong
Liu
L
The National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.
Xiaoai
Zhang
Z
Yuanbo
Cui
C
Jiachang
Yue
Y
Zhiyong
Luo
L
Peidong
Jiang
J
D005456:Fluorescent Dyes; D006863:Hydrogen-Ion Concentration; D006180:Proton-Translocating ATPases / Q000378:metabolism*; D011522:Protons; D012247:Rhodospirillum rubrum
D005456:Fluorescent Dyes; D011522:Protons; D006180:Proton-Translocating ATPases