The Tyr(2)(+) decay kinetics have been analyzed by using time-resolved
EPR to determine the half-time of each S-i --> S-(i+l) transition in
the O-2-evolving complex of spinach thylakoids under physiological con
ditions. Using dark-adapted thylakoids and appropriate single-turnover
flash sequences, we wen able to detect the signal IIvf kinetics of th
e Tyr(z)(+) S-0 --> Tyr(z)(+) S-1, Tyr(z)(+) S-1 --> Tyr(z) S-2, Tyr(z
)(+) S-2 --> Tyr(z) S-3, and Tyr(z)(+) S-3 --> (S-4) --> Tyr(z) S-0 tr
ansitions. To correct for damping of the S state synchronization durin
g the Rash sequence, the Kok parameters were estimated by measuring th
e oxygen flash pattern in situ using nitroxide-based EPR oximetry. Fol
lowing deconvolution of the individual S slate contributions, the sign
al IIvf decay kinetics yield the following half-times for the S state
transitions: S-0 --> S-1 in 40-60 mu s, S-1 --> S-2 in 85 mu s, S-2 --
> S-3 in 140 mu s, and S-3 --> (S-4) --> S-0 in 750 mu s. Preliminary
results with detergent-solubilized PSII membranes suggest that the S-3
--> S-0 transition at least is slowed by a factor of similar to 2 in
this system. Ramifications of these half-times in terms of electron tr
ansfer events on the donor site of PSII are discussed.