Light activates rotations of bacteriorhodopsin in the purple membrane.

To investigate how a photoactivated chromophore drives the proton pump mechanism of bacteriorhodopsin, we have observed how the chromophore rotates during the photocyle. To do this, we examined the dichroism induced in aqueous suspensions of purple membrane fragments by flashes of linearly polarized light. We find that the flash stimulates both the photocycling chromophores and their noncycling neighbors to undergo large (greater than 10 degrees - 20 degrees) rotations within the membrane during the photocycle, and that these two chromophore populations undergo distinctly different sequences of rotations. All these rotations could be eliminated by glutaraldehyde fixation as well as by embedding unfixed fragments in polyacrylamide or agarose gels. Thus, in these immbolizing preparations the chromophore can photocycle without rotating inside a bacteriorhodopsin monomer by more than our detection limit of 2 degrees - 5 degrees. The large rotations we observed in aqueous suspensions of purple membranes were probably due to rotations of entire protein monomers. The process by which a photocycling monomer causes its noncycling neighbors to rotate may help explain the highly cooperative behavior bacteriorhodopsin exhibits when it is aggregated into crystalline arrays of trimers.