PHOTOACTIVITY OF TRANSIENT INTERMEDIATES IN THE PATHWAY FROM THE RED-ABSORBING TO THE FAR-RED-ABSORBING FORM OF Avena PHYTOCHROME AS OBSERVED BY A DOUBLE-FLASH TRANSIENT-SPECTRUM ANALYZER

Abstract— Phototransformation from the red-absorbing form of phytochrome (Pr) to the far-red-absorbing form occurs via at least two reaction stages. We report here on the photoactivity of the intermediates present during these two stages, as detected with 114/118-kdalton Avena phytochrome by a transient spectrum analyzer with two actinic lasers of microsecond duration. Flash activation of intermediates present during the first stage results in their photoconversion back to Pr within the time resolution provided by the analyzer, which is 10 μs. Since these intermediates are present within 60 ns of excitation of Pr (Cordonnier et al. , 1981), a single flash of ms duration as used here should yield a photostationary equilibrium between them and Pr. The proportion of Pr converted to intermediates by a single saturating flash is estimated to be about 30%. Flash activation of intermediates present during the second stage converts them to another transient intermediate stage, which decays thermally to Pr within 2 ms.