The effect of illumination at 5 K of photosystem II in different S-states was investigated with EPR spectroscopy. Two split radical EPR signals around g approximate to 2.0 were observed from samples given 0 and 3 flashes, respectively. The signal from the 0-flash sample was narrow, with a width of similar to80 G, in which the low-field peak can be distinguished. This signal oscillated with the S, state in the sample. The signal from the 3-flash sample was broad, with a symmetric shape of similar to160 G width from peak to trough. This signal varied with the concentration of the So state in the sample. Both signals are assigned to arise from the donor side of PSII Both signals relaxed fast, were formed within 10 ms after a flash, and decayed with half-times at 5 K of 3-4 min. The signal in the So state closely resembles split radical signals, originating from magnetic interaction between Y-Z(.) and the S-2 state, that were first observed in Ca2+-depleted photosystem II samples. Therefore, we assign this signal to Y-Z(.) in magnetic interaction with the S-0 state, Yz(.)S(0). The other signal is assigned to the magnetic interaction between Y-Z(.) and the S-1 state, (YZS1)-S-.. An important implication is that Y-Z can be oxidized at 5 K in the S-0 and S-1 states. Oxidation of Y-Z involves deprotonation of the tyrosine. This is restricted at 5 K, and we therefore suggest that the phenolic proton of Y-Z is involved in a low-barrier hydrogen bond. This is an unusually short hydrogen bond in which proton movement at very low temperatures can occur.
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