Kato-Yamada Y, Noji H, Yasuda R, Kinosita K, Yoshida M
Research Laboratory of Resources Utilization, Tokyo Institute of Technology, 4259 Nagatsuta, Yokohama, 226-8503, Japan.
J Biol Chem. 1998 Jul 31;273(31):19375-7. doi: 10.1074/jbc.273.31.19375.
Rotation of the epsilon subunit in F1-ATPase from thermophilic Bacillus strain PS3 (TF1) was observed under a fluorescence microscope by the method used for observation of the gamma subunit rotation (Noji, H., Yasuda, R., Yoshida, M., and Kinosita, K., Jr. (1997) Nature 386, 299-302). The alpha3 beta3 gamma epsilon complex of TF1 was fixed to a solid surface, and fluorescently labeled actin filament was attached to the epsilon subunit through biotin-streptavidin. In the presence of ATP, the filament attached to epsilon subunit rotated in a unidirection. The direction of the rotation was the same as that observed for the gamma subunit. The rotational velocity was slightly slower than the filament attached to the gamma subunit, probably due to the experimental setup used. Thus, as suggested from biochemical studies (Aggeler, R., Ogilvie, I. , and Capaldi, R. A. (1997) J. Biol. Chem. 272, 19621-19624), the epsilon subunit rotates with the gamma subunit in F1-ATPase during catalysis.
通过用于观察γ亚基旋转的方法（野地，H.，安田，R.，吉田，M.，和木下，K.，Jr.（1997年）《自然》386，299 - 302），在荧光显微镜下观察了嗜热芽孢杆菌PS3菌株（TF1）的F1 - ATP酶中ε亚基的旋转。TF1的α3β3γε复合物固定在固体表面，荧光标记的肌动蛋白丝通过生物素 - 链霉亲和素连接到ε亚基上。在ATP存在的情况下，连接到ε亚基的丝单向旋转。旋转方向与观察到的γ亚基的旋转方向相同。旋转速度略慢于连接到γ亚基的丝，这可能是由于所使用的实验设置。因此，正如生化研究（阿格勒，R.，奥格尔维，I.，和卡帕尔迪，R. A.（1997年）《生物化学杂志》272，19621 - 19624）所表明的，在催化过程中，F1 - ATP酶中的ε亚基与γ亚基一起旋转。
