Determination of the primary charge separation rate in isolated photosystem II reaction centers with 500-fs time resolution.
We have measured directly the rate of formation of the oxidized chlorophyll a electron donor (P680(+)) and the reduced electron acceptor pheophytin a(-) (Pheoa(-)) following excitation of isolated spinach photosystem II reaction centers at 4 degrees C. The reaction-center complex consists of D(1), D(2), and cytochrome b-559 proteins and was prepared by a procedure that stabilizes the protein complex. Transient absorption difference spectra were measured from 440 to 850 nm as a function of time with 500-fs resolution following 610-nm laser excitation. The formation of P680(+)-Pheoa(-) is indicated by the appearance of a band due to P680(+) at 820 nm and corresponding absorbance changes at 505 and 540 nm due to formation of Pheoa(-). The appearance of the 820-nm band is monoexponential with tau = 3.0 +/- 0.6 ps. The time constant for decay of (1*)P680, the lowest excited singlet state of P680, monitored at 650 nm, is tau = 2.6 +/- 0.6 ps and agrees with that of the appearance of P680(+) within experimental error. Treatment of the photosystem II reaction centers with sodium dithionite and methyl viologen followed by exposure to laser excitation, conditions known to result in accumulation of Pheoa(-), results in formation of a transient absorption spectrum due to (1*)P680. We find no evidence for an electron acceptor that precedes the formation of Pheoa(-).
Chemistry Division, Argonne National Laboratory, Argonne, IL 60439.
M R
MR
Wasielewski
D G
DG
Johnson
M
M
Seibert
Govindjee