[Light-induced changes in pH and buffer capacity in reaction center preparations of Rhodopseudomonas spheroides]

In experiments with reaction centers (RC) preparations of Rps. sphaeroides, strain 1760-1, pH of the suspension medium was found to change under continuous light excitation. The concentration of hydrogen ions starts to decrease fastly (t1/2 = 2--4 s) after the light is on, whereupon it increases slowly, to a steady-state level, within t1/2 = 5--7 min. During the fast phase 0.3 microM H+/1 microM of RC are absorbed and 7--8 microM H+/1 microM of RC are released under the slow phase. The fast phase is suppressed by ortho-phenantroline (10 mM)--an inhibitor of electron transfer from the primary (Fe-quinone complex) to the secondary (quinone) electron acceptors. Glutaraldehyde which is known to modify a protein structure of the RC inhibits the slow phase. The obtained data suggest that fast light-induced pH changes are due to a H+ uptake by the secondary quinoid anion-radical products; slow pH changes are ascribed to structural changes within the RC complex, which lead to a pK shift and/or changes in the amount of dissociating groups on the protein interfacial surface. The conclusion was supported by an observation of light-induced changes in buffer capacity of an RC suspension and by a light-induced increase in the amount of SH-groups titrated by 5.5-dithio-bis(2 nitro)-benzoic acid. The role of conformational dynamics of photoactive pigment--protein complexes in the functions of the RC is discussed.