Temperature control of the motility of actin filaments interacting with myosin molecules using an electrically conductive glass in the presence of direct current

• Published in: Archives of biochemistry and biophysics Vol. 586; pp. 51 - 56
• Main Authors: Wada, Reito, Sato, Daisuke, Nakamura, Takao, Hatori, Kuniyuki
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15.11.2015
• Subjects: Actin Cytoskeleton - chemistry; Actin Cytoskeleton - metabolism; Animals; Biophysical Phenomena; Electric Conductivity; Glass; In Vitro Techniques; Indium tin oxide; Molecular Motor Proteins - chemistry; Molecular Motor Proteins - metabolism; Motility assay; Motion; Motor protein; Myosin Subfragments - chemistry; Myosin Subfragments - metabolism; Rabbits; Temperature; Temperature dependence; Tin Compounds; Velocity distribution; Temperature dependence; Motility assay; Motor protein; Indium tin oxide; ITO; HMM; Velocity distribution; ATP
• ISSN: 0003-9861; 1096-0384
• DOI: 10.1016/j.abb.2015.10.001

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. •Surface current modulates temperature of ITO-glass for fluorescence microscopy.•Current flow did not affect filament direction in actomyosin motility assays.•Filament velocity distribution has a broad range that is temperature-dependent.