Statistics of reversible bond dynamics observed in force-clamp spectroscopy

TL;DR

This paper analyzes two-state trajectories from force-clamp spectroscopy to understand reversible bond dynamics, comparing event and cycle counting methods, and exploring dynamic disorder and Markovian assumptions in complex systems.

Contribution

It introduces a detailed analysis framework for FCS data, including cycle-counting and dynamic disorder considerations, enhancing understanding of force-dependent kinetics.

Findings

Cycle-counting can be advantageous when transitions are hard to resolve.

Analysis of dynamic disorder provides insights into force-dependent transition rates.

The study discusses the applicability of Markovian models to complex two-state systems.

Abstract

We present a detailed analysis of two-state trajectories obtained from force-clamp spectroscopy (FCS) of reversibly bonded systems. FCS offers the unique possibility to vary the equilibrium constant in two-state kinetics, for instance the unfolding and refolding of biomolecules, over many orders of magnitude due to the force dependency of the respective rates. We discuss two different kinds of counting statistics, the event-counting usually employed in the statistical analysis of two-state kinetics and additionally the so-called cycle-counting. While in the former case all transitions are counted, cycle-counting means that we focus on one type of transitions. This might be advantageous in particular if the equilibrium constant is much larger or much smaller than unity because in these situations the temporal resolution of the experimental setup might not allow to capture all transitions of an event-counting analysis. We discuss how an analysis of FCS data for complex systems exhibiting dynamic disorder might be performed yielding information about the detailed force-dependence of the transition rates and about the time scale of the dynamic disorder. In addition, the question as to which extent the kinetic scheme can be viewed as a Markovian two-state model is discussed.