Reaction centre photochemistry in cyanide-treated photosystem II

EPR was used to study the triplet state of chlorophyll generated by radical pair recombination in the photosystem II (PSII) reaction centre. The spin state of the non-haem Fe2+ was varied using the CN--binding method (Y. Sanakis, V. Petrouleas, B.A. Diner, Biochemistry 33 (1994) 9922-9928) and the redox state of the quinone acceptor (QA) was changed from semi-reduced to fully reduced (F.J.E. van Mieghem, W. Nitschke, P. Mathis, A.W. Rutherford, Biochim. Biophys. Acta 977 (1989) 207-214). It was found that the triplet was not detectable using continuous wave EPR when QA- was present irrespective of the spin-state of the Fe2+. It was also found that the triplet state became detectable by EPR when the semiquinone was removed (by reduction to the quinol) and that the triplet observed was not influenced by the spin state of the Fe2+. Since it is known from earlier work that the EPR detection of the triplet reflects a change in the triplet lifetime, it is concluded that the redox state of the quinone determines the triplet lifetime (at least in terms of its detectability by continuous wave EPR) and that the magnetic state of the iron, (through the weakly exchange-coupled QA- Fe2+ complex) is not a determining factor. In addition, we looked for polarisation transfer from the radical pair to QA- in PSII where the Fe2+ was low spin. Such polarisation is seen in bacterial reaction centres under comparable conditions. In PSII, however, we were unable to find evidence for such polarisation of the semiquinone. It is suggested that both the short triplet lifetime in the presence of QA- and the lack of polarised QA- might be explained in terms of the electron transfer mechanism for triplet quenching involving the semiquinone which was proposed previously (F.J.E. van Mieghem, K. Brettel, B. Hillmann, A. Kamlowski, A.W. Rutherford, E. Schlodder, Biochemistry 34 (1995) 4798-4813). It is suggested that this mechanism may occur in PSII (but not in purple bacterial reaction centres) due the triplet-bearing chlorophyll being adjacent to the pheophytin at low temperature as suggested from structural studies (F.J.E. van Mieghem, K. Satoh, A.W. Rutherford, Biochim. Biophys. Acta 1058 (1992) 379-385).