Evidence for protein dielectric relaxations in reaction centers associated with the primary charge separation detected from Rhodospirillum rubrum chromatophores by combined photovoltage and absorption measurements in the 1-15 ns time range.

Abstract

Fast photovoltage measurements in Rhodospirillum rubrum chromatophores in the nanosecond time range, escorted by time-resolved absorption measurements, are described. Under reducing conditions, the photovoltage decayed significantly faster than the spectroscopically detected charge recombination of the radical pair P(+)H(A)(-). This indicates the occurrence of considerable dielectric relaxations. Our data and data from the literature were analyzed by means of a reaction scheme consisting of three states, namely, A, P, and P(+)H(A)(-). A time-dependent DeltaG(t) was introduced by assuming a time-dependent rate constant of the back-reaction, k(-1)(t). With the exception of the latter rate constant, all other parameters of the model are reliably known within narrow limits. This allowed us to distinguish between the three cases assumed for DeltaG degrees (t): (1)DeltaG degrees (t) = constant; (2)DeltaG degrees (t) as published by Peloquin et al. [Peloquin, J. M., Williams, J. C., Lin, X. M., Alden, R. G., Taguchi, A. K. W., Allen, J. P., and Woodbury, N. W. (1994) Biochemistry 33, 8089-8100]; and a (3)DeltaG degrees (t) that fits the present data. The assumption that (1)DeltaG degrees (t) = constant is incompatible with our photovoltage data, and (2)DeltaG degrees (t) is incompatible with the constraint that the ratio of fluorescence yields in the closed and open state is F(m)/F(o) approximately 2. We specify a (3)DeltaG degrees (t) that should be valid for photosynthetic reaction centers in vivo. Furthermore, the overall kinetics of the electric relaxation, e(t), in response to the primary charge separation were determined.