The identification of a unique intermediate in biological water oxidation establishes the water binding mechanism in the S2 to S3 state transition.

Flash-induced chlorophyll fluorescence kinetics from photosystem II in thylakoids from the dark-grown wild type and two site-directed mutants of the D1 protein His190 residue (D1-H190) in Chlamydomonas reinhardtii have been characterized. Induction of the chlorophyll fluorescence on the first flash, reflecting electron transport from YZ to P680(+), exhibited a strong pH dependence with a pK of 7.6 in the dark-grown wild type which lacks the Mn cluster. The chlorophyll fluorescence decay, measured in the presence of DCMU, which reflects recombination between QA- and YZox, was also pH-dependent with a similar pK of 7.5. These results indicate participation by the same base, which is suggested to be D1-H190, in oxidation and reduction of YZ in forward electron transfer and recombination pathways, respectively. This hypothesis was tested in the D1-H190 mutants. Induction of chlorophyll fluorescence in these H190 mutants has been observed to be inefficient due to slow electron transfer from YZ to P680(+) [Roffey, R. A., et al. (1994) Biochim. Biophys. Acta 1185, 257-270]. We show that this reaction is pH-dependent, with a pK of 8. 1, and at pH >/=9, the fluorescence induction is efficient in the H190 mutants, suggesting direct titration of YZ. The efficient oxidation of YZ ( approximately 70% at pH 9.0) at high pH was confirmed by kinetic EPR measurements. In contrast to the wild type, the H190 mutants show little or no observable fluorescence decay. Our data suggest that H190 is an essential component in the electron transfer reactions in photosystem II and acts as a proton acceptor upon YZ oxidation. In the H190 mutants, this reaction is inefficient and YZ oxidation only occurs at elevated pHs when YZ itself probably is deprotonated. We also propose that H190 is able to return a proton to YZox during electron recombination from QA- in a reaction which does not take place in the D1-H190 mutants.

No abstract

Photosystem II (PSII), the thylakoid membrane enzyme which uses sunlight to oxidize water to molecular oxygen, holds many organic and inorganic redox cofactors participating in the electron transfer reactions. Among them, two tyrosine residues, Tyr-Z and Tyr-D are found on the oxidizing side of PSII. Both tyrosines demonstrate similar spectroscopic features while their kinetic characteristics are quite different. Tyr-Z, which is bound to the D1 core protein, acts as an intermediate in electron transfer between the primary donor, P(680) and the CaMn₄ cluster. In contrast, Tyr-D, which is bound to the D2 core protein, does not participate in linear electron transfer in PSII and stays fully oxidized during PSII function. The phenolic oxygens on both tyrosines form well-defined hydrogen bonds to nearby histidine residues, His(Z) and His(D) respectively. These hydrogen bonds allow swift and almost activation less movement of the proton between respective tyrosine and histidine. This proton movement is critical and the phenolic proton from the tyrosine is thought to toggle between the tyrosine and the histidine in the hydrogen bond. It is found towards the tyrosine when this is reduced and towards the histidine when the tyrosine is oxidized. The proton movement occurs at both room temperature and ultra low temperature and is sensitive to the pH. Essentially it has been found that when the pH is below the pK(a) for respective histidine the function of the tyrosine is slowed down or, at ultra low temperature, halted. This has important consequences for the function also of the CaMn₄ complex and the protonation reactions as the critical Tyr-His hydrogen bond also steer a multitude of reactions at the CaMn₄ cluster. This review deals with the discovery and functional assignments of the two tyrosines. The pH dependent phenomena involved in oxidation and reduction of respective tyrosine is covered in detail. This article is part of a Special Issue entitled: Photosystem II.