How Does F(1)-ATPase Generate Torque?: Analysis From Cryo-Electron Microscopy and Rotational Catalysis of Thermophilic F(1)

The F(1)-ATPase is a rotary motor fueled by ATP hydrolysis. Its rotational dynamics have been well characterized using single-molecule rotation assays. While F(1)-ATPases from various species have been studied using rotation assays, the standard model for single-molecule studies has been the F(1)-ATPase from thermophilic Bacillus sp. PS3, named TF(1). Single-molecule studies of TF(1) have revealed fundamental features of the F(1)-ATPase, such as the principal stoichiometry of chemo-mechanical coupling (hydrolysis of 3 ATP per turn), torque (approximately 40 pN·nm), and work per hydrolysis reaction (80 pN·nm = 48 kJ/mol), which is nearly equivalent to the free energy of ATP hydrolysis. Rotation assays have also revealed that TF(1) exhibits two stable conformational states during turn: a binding dwell state and a catalytic dwell state. Although many structures of F(1) have been reported, most of them represent the catalytic dwell state or its related states, and the structure of the binding dwell state remained unknown. A recent cryo-EM study on TF(1) revealed the structure of the binding dwell state, providing insights into how F(1) generates torque coupled to ATP hydrolysis. In this review, we discuss the torque generation mechanism of F(1) based on the structure of the binding dwell state and single-molecule studies.