Domain-domain interactions in Filamin A (16-23) impose a hierarchy of unfolding forces

TL;DR

This study uses single molecule force spectroscopy to reveal how domain interactions in Filamin A (16-23) create a hierarchical mechanical response, enabling it to withstand diverse cellular forces.

Contribution

It demonstrates the existence of a force-dependent hierarchy of domain rupture and unfolding in Filamin A (16-23), highlighting its complex mechanical behavior compared to linear forms.

Findings

Filamin A 16-23 exhibits a broad distribution of rupture forces and unfolding times.

A mechanical hierarchy exists where high-force rupture enables low-force unfolding.

Filamin A 16-23 is more compliant under 100 pN than the linear FLNa 1-8.

Abstract

The quaternary structure of Filamin A (FLNa) 16-23 was recently shown to exhibit multiple domain-domain interactions that lead to a propeller-like construction. Here we present single molecule force spectroscopy experiments to show a wide variety of mechanical responses of this molecule and compare it with its linear counterpart FLNa 1-8. The compact structure of FLNa 16-23 leads to a broad distribution of rupture forces and end-to-end lengths in the force-extension mode and multiple unraveling timescales in the force-clamp mode. Moreover, a subset of force-extension trajectories reveals a mechanical hierarchy in which the rupture of domain-domain interactions at high forces (200 pN) liberates the unfolding of individual domains at low forces (100 pN). This mechanism may also explain the order of magnitude difference in the rates of the biexponential fits to the distribution of unfolding dwell times under force-clamp. Overall, FLNa 16-23 under a force of 100 pN is more compliant than the linear FLNa 1-8. Since a physiological role of FLNa is to crosslink actin filaments, this range of responses allows it to accommodate a broad spectrum of forces exerted by the cell and its environment.