Kinetic coupling of phosphate release, force generation and rate-limiting steps in the cross-bridge cycle

A basic goal in muscle research is to understand how the cyclic ATPase activity of cross-bridges is converted into mechanical force. A direct approach to study the chemo-mechanical coupling between Pi release and the force-generating step is provided by the kinetics of force response induced by a rapid change in [Pi]. Classical studies on fibres using caged-Pi discovered that rapid increases in [Pi] induce fast force decays dependent on final [Pi] whose kinetics were interpreted to probe a fast force-generating step prior to Pi release. However, this hypothesis was called into question by studies on skeletal and cardiac myofibrils subjected to Pi jumps in both directions (increases and decreases in [Pi]) which revealed that rapid decreases in [Pi] trigger force rises with slow kinetics, similar to those of calcium-induced force development and mechanically-induced force redevelopment at the same [Pi]. A possible explanation for this discrepancy came from imaging of individual sarcomeres in cardiac myofibrils, showing that the fast force decay upon increase in [Pi] results from so-called sarcomere 'give'. The slow force rise upon decrease in [Pi] was found to better reflect overall sarcomeres cross-bridge kinetics and its [Pi] dependence, suggesting that the force generation coupled to Pi release cannot be separated from the rate-limiting transition. The reasons for the different conclusions achieved in fibre and myofibril studies are re-examined as the recent findings on cardiac myofibrils have fundamental consequences for the coupling between Pi release, rate-limiting steps and force generation. The implications from Pi-induced force kinetics of myofibrils are discussed in combination with historical and recent models of the cross-bridge cycle.