Detachment, Futile Cycling and Nucleotide Pocket Collapse in Myosin-V Stepping

TL;DR

This study develops and compares mechanochemical models of myosin-V stepping, identifying key mechanisms that reproduce experimental velocity and run-length trends, including nucleotide-dependent detachment and futile cycling.

Contribution

The paper introduces comprehensive reaction network models that accurately replicate all observed experimental trends in myosin-V motility, highlighting the roles of nucleotide-dependent detachment and futile cycling.

Findings

Models with [ADP]-dependent detachment reproduce all trends.

Futile cycling and nucleotide pocket collapse are crucial mechanisms.

Kinetic transition rates favor futile cycling as the key process.

Abstract

Myosin-V is a highly processive dimeric protein that walks with 36nm steps along actin tracks, powered by coordinated ATP hydrolysis reactions in the two myosin heads. No previous theoretical models of the myosin-V walk reproduce all the observed trends of velocity and run-length with [ADP], [ATP] and external forcing. In particular, a result that has eluded all theoretical studies based upon rigorous physical chemistry is that run length decreases with both increasing [ADP] and [ATP]. We systematically analyse which mechanisms in existing models reproduce which experimental trends and use this information to guide the development of models that can reproduce them all. We formulate models as reaction networks between distinct mechanochemical states with energetically determined transition rates. For each network architecture, we compare predictions for velocity and run length to a subset of experimentally measured values, and fit unknown parameters using a bespoke MCSA optimization routine. Finally we determine which experimental trends are replicated by the best-fit model for each architecture. Only two models capture them all: one involving [ADP]-dependent mechanical detachment, and another including [ADP]-dependent futile cycling and nucleotide pocket collapse. Comparing model-predicted and experimentally observed kinetic transition rates favors the latter.