The association of biomolecules is the elementary event of communication in biology. Most mechanistic information of how the interactions between binding partners form or break is, however, hidden in the transition paths, the very short parts of the molecular trajectories from the encounter of the two molecules to the formation of a stable complex. Here we use single-molecule spectroscopy to measure the transition path times for the association of two intrinsically disordered proteins that form a folded dimer upon binding. The results reveal the formation of a metastable encounter complex that is electrostatically favored and transits to the final bound state within tens of microseconds. Such measurements thus open a new window into the microscopic events governing biomolecular interactions.

In this study we investigate the effect of shear force on lipid membranes induced by external fluid flow. We use giant unilamellar vesicles (GUVs) as simple cell models and chose a ternary lipid mixture that exhibits liquid-ordered and liquid-disordered domains. These domains are stained with different dyes to allow visualization of changes within the membrane after the application of flow. A microfluidic device served as a valuable platform to immobilize the vesicles and apply shear forces of a defined strength. Moreover, integration of valves allowed us to stop the flow instantaneously and visualize the relaxing domain patterns by means of high-resolution fluorescence microscopy. We observed the formation of transient, non-deterministic patterns of the formerly round domains during application of flow. When the flow is stopped, round domains are formed again on a time scale of ms to s. At longer time scales of several seconds to minutes, the domains fuse into larger domains until they reach equilibrium. These processes are accelerated with increasing temperature and vesicles with budding domains do not fuse unless the temperature is elevated. Our results show the strong effect of the flow on the lipid membrane and we believe that this phenomenon plays a crucial role in the processes of mechanotransduction in living cells.

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