title: Flash kinetic study of the last steps in the photoinduced reaction cycle of bacteriorhodopsin.
abstract: The reaction cycle of light adapted bacteriorhodopsin (BR) in aqueous purple membrane suspensions was studied by laser flash photolysis at different temperatures (2--49 degrees C) and pH values (3--10). The activation energy for several reaction steps was determined at pH 7.6. The kinetics of O-bacteriorhodopsin (one of the last intermediates in the cycle) were analyzed in some detail and it was found that the simple consecutive reaction scheme M-BR leads to O-BR leads to BR may explain the kinetics of O-bacteriorhodopsin as measured at 680 nm. Since the pH change in neutral aqueous suspensions of purple membrane follows similar kinetics as O-bacteriorhodopsin it is suggested that protons are released during the reaction M-BR leads to O-BR and taken up again during the reacton O-BR leads to BR. Another long-lived intermediate, which absorbs to a greater extent than bacteriorhodopsin at 570 nm and less than bacteriorhodopsin at 420 nm, was identified with the strongly fluorescing species, pseudo- or P-bacteriorhodopsin. The decay of P-bacteriorhodopsin in bacteriohodopsin had an activation energy of only approx. 1.2 kcal/mol, which suggests that the last step of the photocycle is a relaxation around a single bond. At pH 9--10, the simple first-order kinetics of all the intermediates were changed into a kinetics consisting of two first-order decays. This change of kinetics was accompanied by a drastic decrease in the rotational diffusion relaxation time. To explain the results obtained in this work and those of others, a model involving proton uptake and release by the Schiff base nitrogen combined with an isomerization reaction is finally proposed.
mesh_terms: D001436:Bacteriorhodopsins / Q000378:metabolism*; D002338:Carotenoids / Q000378:metabolism*; D006217:Halobacterium / Q000378:metabolism*; D007700:Kinetics; D007834:Lasers; D008027:Light; D010782:Photolysis; D010788:Photosynthesis; D013696:Temperature; D013816:Thermodynamics
chemical_list: D002338:Carotenoids; D001436:Bacteriorhodopsins