Bacteriorhodopsin photocycle kinetics analyzed by the maximum entropy method

Abstract

A maximum entropy method (MEM) was developed for the study of the bacteriorhodopsin photocycle kinetics. The method can be applied directly to experimental kinetic absorption data without any assumption for the number of the intermediate states taking part in the photocycle. Though this method does not give a specific kinetics, its result is very useful for selection between possible photocycle kinetics. Using simulated data, it is shown that MEM gives correct results for the number of the intermediate states and the amplitude distributions around the characteristic lifetimes. Analyzing experimental absorption data at five different wavelengths, MEM gives seven or eight characteristic lifetimes, which means that at least so many distinct intermediate states exist during the photocycle. Many possible photocycle kinetic models were studied and compared with the MEM result. The best agreement was found with a branching photocycle model of eight intermediate states ( K, L, M 1, M 2, M 3, M 4, N, O). The branching occurs at the L intermediate state ( M 1 and M 2 being in one branch and M 3 and M 4 in the other branch), but at high pH it occurs already at the K state.