Abstract
Cell adhesion complexes (CACs), which are activated by ligand binding, play key roles in many cellular functions ranging from cell cycle regulation to mediation of cell extracellular matrix adhesion. Inspired by single molecule pulling experiments using atomic force spectroscopy on leukocyte function-associated antigen-1 (LFA-1), expressed in T-cells, bound to intercellular adhesion molecules (ICAM), we performed constant loading rate (r(f)) and constant force (F) simulations using the self-organized polymer model to describe the mechanism of ligand rupture from CACs. The simulations reproduce the major experimental finding on the kinetics of the rupture process, namely, the dependence of the most probable rupture forces (f*s) on ln r(f) (r(f) is the loading rate) exhibits two distinct linear regimes. The first, at low r(f), has a shallow slope, whereas the slope at high r(f) is much larger, especially for a LFA-1/ICAM-1 complex with the transition between the two occurring over a narrow r(f) range. Locations of the two transition states (TSs) extracted from the simulations show an abrupt change from a high value at low r(f) or constant force, F, to a low value at high r(f) or F. This unusual behavior in which the CACs switch from one brittle (TS position is a constant over a range of forces) state to another brittle state is not found in forced-rupture in other protein complexes. We explain this novel behavior by constructing the free energy profiles, F(Λ)s, as a function of a collective reaction coordinate (Λ), involving many key charged residues and a critical metal ion (Mg(2+)). The TS positions in F(Λ), which quantitatively agree with the parameters extracted using the Bell-Evans model, change abruptly at a critical force, demonstrating that it, rather than the molecular extension, is a good reaction coordinate. Our combined analyses using simulations performed in both the pulling modes (constant r(f) and F) reveal a new mechanism for the two loading regimes observed in the rupture kinetics in CACs.