Controlling the Motility of ATP-Driven Molecular Motors Using High Hydrostatic Pressure
High-pressure microscopy is a powerful technique for visualizing the effects of hydrostatic pressure on research targets. It can be used for monitoring the pressure-induced changes in the structure and function of molecular machines in vitro and in vivo. This chapter focuses on the use of high-pressure microscopy to measure the dynamic properties of molecular machines. We describe a high-pressure microscope that is optimized both for the best image formation and for stability under high hydrostatic pressure. The developed system allows us to visualize the motility of ATP-driven molecular motors under high pressure. The techniques described could be extended to study the detailed mechanism by which molecular machines work efficiently in collaboration with water molecules.
KeywordsHigh-pressure microscopy Molecular motors Kinesin F1-ATPase Single-molecule measurement
We would like to thank Yoshifumi Kimura for developing a prototype of the high-pressure chamber for optical microscopy, Daichi Okuno and Hiroyuki Noji for the measurement of F1-ATPase, and Eiro Muneyuki, Shoichi Toyabe, Shou Furuike, Masahide Terazima, Yoshie Harada, and Akitoshi Seiyama for discussion. This work was supported by Grants-in-Aid for Scientific Research on the Innovative Area “Water plays the main role in ATP energy transfer, Group Leader; Prof. Makoto Suzuki” (Nos. 21118511 and 23118710), Grants-in-Aid for Scientific Research from MEXT (Nos. 16 K04908 and 17H05880), and the Takeda Science Foundation, Research Foundation for Opto-Science and Technology, Shimadzu Science Foundation, and Nakatani Foundation for Advancement of Measuring Technologies in Biomedical Engineering.
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