The relative weight of the slowly decaying M intermediate of the photocycle of bacteriorhodopsin increases upon increasing the energy density of the short (10 ns) actinic laser pulse. Moreover, when a pre-exciting flash is applied to the BR sample, the absolute amplitude of the Ms is higher in the signal induced by a second flash, applied with a delay from 100 microseconds to 100 ms. These facts together prove that either the leftover BR ground-state population becomes different due to the pre-excitation, or there is a cooperative interaction between the BR molecules.
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Tokaji et al. (1991) studied this question.
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