Inter-subunit rotation and elastic power transmission in F 0F 1-ATPase

Abstract

ATP synthase (F-ATPase) produces ATP at the expense of ion-motive force or vice versa. It is composed from two motor/generators, the ATPase (F 1) and the ion translocator (F 0), which both are rotary steppers. They are mechanically coupled by 360° rotary motion of subunits against each other. The rotor, subunits γϵ c 10–14, moves against the stator, (αβ) 3δ ab 2. The enzyme copes with symmetry mismatch (C 3 versus C 10–14) between its two motors, and it operates robustly in chimeric constructs or with drastically modified subunits. We scrutinized whether an elastic power transmission accounts for these properties. We used the curvature of fluorescent actin filaments, attached to the rotating c ring, as a spring balance (flexural rigidity of 8·10 −26 N m 2) to gauge the angular profile of the output torque at F 0 during ATP hydrolysis by F 1. The large average output torque (56 pN nm) proved the absence of any slip. Angular variations of the torque were small, so that the output free energy of the loaded enzyme decayed almost linearly over the angular reaction coordinate. Considering the three-fold stepping and high activation barrier (>40 kJ/mol) of the driving motor (F 1) itself, the rather constant output torque seen by F 0 implied a soft elastic power transmission between F 1 and F 0. It is considered as essential, not only for the robust operation of this ubiquitous enzyme under symmetry mismatch, but also for a high turnover rate under load of the two counteracting and stepping motors/generators.