Flash Inactivation of Oxygen Evolution: IDENTIFICATION OF S(2) AS THE TARGET OF INACTIVATION BY TRIS

Brief saturating light flashes were used to probe the mechanism of inactivation of O(2) evolution by Tris in chloroplasts. Maximum inactivation with a single flash and an oscillation with period of four on subsequent flashes was observed. Analyses of the oscillations suggested that only the charge-collecting O(2)-evolving catalyst of photosystem II (S(2)-state) was a target of inactivation by Tris. This conclusion was supported by the following observations: (a) hydroxylamine preequilibration caused a three-flash delay in the inactivation pattern; (b) the lifetimes of the Tris-inactivable and S(2)-states were similar; and (c) reagents accelerating S(2) deactivation decreased the lifetime of the inactivable state. Inactivation proved to be moderated by F, the precursor of Signal II(s), as shown by a one flash delay with chloroplasts having high abundance of F. Evidence was obtained for cooperativity effects in inactivation and NH(3) was shown to be a competitive inhibitor of the Tris-induced inactivation. S(2)-dependent inactivation was inhibited by glutaraldehyde fixation of chloroplasts, possibly suggesting that inactivation proceeds via conformational changes of the S(2)-state.