Site-directed mutations at D1-His198 and D1-Thr179 of photosystem II in Synechocystis sp. PCC 6803: deciphering the spectral properties of the PSII reaction centre

Abstract

Site-directed mutations were constructed in photosystem II of Synechocystis sp. PCC6803 in which the axial ligand, D1-His198, of special pair chlorophyll P _D1 was replaced with Gln and where D1-Thr179, which overlies monomeric chlorophyll Chl _D1 , was replaced with His. The D1-His198Gln mutation produces a 3 nm displacement to the blue of the bleaching minimum in the Soret and in the Qy region of the – absorbance difference spectrum. To a first approximation, the bleaching can be assigned to the low-energy exciton transition of the special pair chlorophylls P _D1 /P _D2 . The D1-Thr179His mutation produces a 2 nm displacement to the red of the bleaching minimum in the Qy region of the ( ³ P– ¹ P) absorbance difference spectrum. Analysis of the flash-induced – and ( ³ P– ¹ P) absorbance difference spectra of both mutants compared with wild-type at 80 K indicate that the cation of the oxidized donor P ⁺ is predominantly localized on the chlorophyll P _D1 of the special pair and that the reaction centre triplet state, produced upon charge recombination from ³ [P ⁺ Pheo ⁻ ], when the primary quinone electron acceptor Q _A is doubly reduced, is primarily localized on Chl _D1 .