Analysis of the reaction coordinate of photosynthetic water oxidation by kinetic measurements of 355 nm absorption changes at different temperatures in photosystem II preparations suspended in either H2O or D2O

Flash-induced absorption changes at 355 nm were measured at different temperatures within the range of 2 degrees C less than or equal to curly theta less than or equal to 25 degrees C in dark-adapted PS II core complexes from spinach [O-2 evolution rate: 1500 +/- 100 mu mol of O-2 (mg of Chi)(-1) h(-1)] that were dissolved either in H2O- or in D2O-containing buffer. Comparative measurements were performed at 20 degrees C in H2O- or D2O-containing suspensions of PS II membrane fragments [O-2 evolution rate: 600 +/- 40 mu mol of O-2 (mg of Chi)(-1) h(-1)]. The results obtained reveal the following: (a) The activation energies of the individual redox steps in the water oxidizing complex (WOC) are dependent on the redox state S-i with E-A(S-1-->S-2) = 14 kJ/mol, E-A(S-2-->S-3) = 35 kJ/mol, and. E-A(S-3-->-->S-0 + O-2) = 21 kJ/mol for curly theta > 11 degrees C, 67 kJ/mol for curly theta < 11 degrees C in PS II core complexes dissolved in H2O; (b) replacement of exchangeable protons by deuterons causes only minor changes (less than or equal to 15%) of the activation energies; and (c) the rate constants of these reactions in PS II fore complexes are characterized by H/D isotope ratios, k(i)(H)/k(i)(D), of 1.6, 2.3, and 1.5 for the transitions S-1-->S-2, S-2-->S3, and S-3-->-->S-0 + O-2, respectively. The corresponding values of PS II membrane fragments are 1.3, 1.3, and 1.4, Based on these results and corresponding E-A data reported in the literature for PS II membrane fragments from spinach [Renger, G., & Hanssum, B. (1992) FEES Lett. 299, 28-32] and PS II particles from the thermophilic cyanobacterium Synechococcus vulcanus Copeland [Koike, II., Hanssum, B., Inoue, Y., & Renger, G. (1987) Biochim. Biophys. Acta 893, 524-533], the reaction coordinate of the redox sequence in the WOC is inferred to be almost invariant to the evolutionary development from cyanobacteria to higher plants. Furthermore, the rather high activation energy of the S-2-->S-3 transition provides evidence fur a significant structural change coupled with this reaction, Implications for the mechanism of photosynthetic water oxidation are discussed.