Active membrane fluctuations with proton pumps

We investigate nonequilibrium membrane fluctuations in giant unilamellar vesicles (GUVs) by reconstituting the light-activated proton pump, bacteriorhodopsin (BR). Once activated, BR is known to form oligomers in model membranes, with an unknown effect on fluctuation amplitude. By using the UV-crosslinkable lipid, 1,2-bis(10,12-tricosadiynoyl)-sn-glycero-3-phosphocholine, we localize BR and prevent light-induced oligomerization. This system allows us to selectively switch on and off active fluctuations and to turn off diffusion (oligomerization) using green~and UV light, respectively. We compare active membrane fluctuations with trapped and freely diffusing BR using an interferometric technique with~sub nanometer and microsecond resolution. Remarkable dependence is found on whether BR is localized or freely diffusing.