Structural snapshots of V/A-ATPase reveal the rotary catalytic mechanism of rotary ATPases

Abstract

V/A-ATPase is a motor protein that shares a common rotary catalytic mechanism with F_oF₁ ATP synthase. When powered by ATP hydrolysis, the V₁ domain rotates the central rotor against the A₃B₃ hexamer, composed of three catalytic AB dimers adopting different conformations (AB_open, AB_semi, and AB_closed). Here, we report the atomic models of 18 catalytic intermediates of the V₁ domain of V/A-ATPase under different reaction conditions, determined by single particle cryo-EM. The models reveal that the rotor does not rotate immediately after binding of ATP to the V₁. Instead, three events proceed simultaneously with the 120˚ rotation of the shaft: hydrolysis of ATP in AB_semi, zipper movement in AB_open by the binding ATP, and unzipper movement in AB_closed with release of both ADP and Pi. This indicates the unidirectional rotation of V/A-ATPase by a ratchet-like mechanism owing to ATP hydrolysis in AB_semi, rather than the power stroke model proposed previously for F₁-ATPase.

The rotary ATPases use a rotary catalytic mechanism to drive transmembrane proton movement powered by ATP hydrolysis. Here, the authors report a collection of V/A-ATPase V₁ domain structures, providing insights into rotary mechanism of the enzyme and potentially other rotary motor proteins driven by ATP hydrolysis.