Wada Reito, Sato Daisuke, Nakamura Takao, Hatori Kuniyuki
Department of Biomedical Information Engineering, Graduate School of Medical Science, Yamagata University, Yamagata 990-9585, Japan.
Department of Bio-Systems Engineering, Graduate School of Science and Engineering, Yamagata University, Yonezawa 992-8510, Japan.
Arch Biochem Biophys. 2015 Nov 15;586:51-6. doi: 10.1016/j.abb.2015.10.001. Epub 2015 Oct 9.
The motility of actin filaments interacting with heavy meromyosin molecules was directly observed on indium tin oxide-coated glass (ITO-glass), over which a surface current flowed. Because the increase in surface current applied to ITO-glass increases the temperature, we focused on the temperature-dependence of the sliding velocity and the effect of the current flow on the orientation of filament motion. Using high precision fluorescence measurements, the displacement vectors of filaments were collected at intervals of 1/30 s. The direction of filament motion was independent to that of current flow up to 0.17 A (7.7 A/m of surface current density); however, the velocity increased by approximately 2-fold when the surface temperature increased from 25 °C to 37 °C. The moving actin filaments exhibited a broader velocity distribution at high temperature than at low temperature. Collectively, these data suggest that using ITO-glass with a surface current to generate a well-controlled temperature change may serve to evaluate temperature-dependent transient responses in protein activity under a microscope, without interference from electrical effects.
在有表面电流流动的氧化铟锡涂层玻璃(ITO玻璃)上,直接观察到了肌动蛋白丝与重酶解肌球蛋白分子相互作用时的运动。由于施加到ITO玻璃上的表面电流增加会使温度升高,我们重点研究了滑动速度的温度依赖性以及电流对丝运动方向的影响。通过高精度荧光测量,每隔1/30秒收集一次丝的位移矢量。在表面电流密度达到0.17 A(7.7 A/m)之前,丝的运动方向与电流方向无关;然而,当表面温度从25℃升高到37℃时,速度增加了约2倍。移动的肌动蛋白丝在高温下比在低温下表现出更宽的速度分布。总体而言,这些数据表明,使用带有表面电流的ITO玻璃来产生可控的温度变化,可能有助于在显微镜下评估蛋白质活性中温度依赖性的瞬态响应,而不受电效应的干扰。