On the primary quantum yields in the bacteriorhodopsin photocycle

Pulsed Nd laser experiments in suspensions of the purple membrane of Halobacterium halobium are carried out at room temperature. At sufficiently high laser intensities, a photostationary mixture of bacteriorhodopsin (BR) and its red-shifted (batho) photoproduct (K) is obtained. The spectra of the first three intermediates in the photocycle are reported. The data yield a value of phi1/phi2=0.40 +/- 0.05 for the ratio of the quantum yields of the forward (phi1) and reverse (phi2) processes, setting an upper limit of approximately 0.4 for the quantum efficiency of the cycle at room temperature. This method is generally available for the determination of phi2 in the case of a photoequilibrium: A in equilibrium B, where B is a short-lived transient and phi1 is known from low intensity measurements. Its potential application is of importance for the study of the photophysics of visual pigments at physiological temperatures