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Influence of KF, DCMU and removal of Ca + on the high-spin EPR signal of the cytochrome b-559 heme Fe(III) ligated by OH - in chloroplasts.
Biochimica et Biophysica Acta 1996 November 12
An EPR signal at g = 6.8 attributed to the cytochrome (Cyt) b-559 heme Fe(III) ligated by OH- (Fiege, R., Schrieber, U., Lubitz, W., Renger, G. and Shuvalov, V.A. (1995) FEBS Lett. 377, 325-329) was studied. This signal is observed in intact chloroplasts when oxidized by 10 mM 2,3-dicyano,5,6-dichloro-p-benzoquinone (DDQ), but not when 5 mM p-benzoquinone is added. Addition of KF (100 mM) or removal of Ca 21 for blocking the water-oxidizing complex considerably decreases the heme Fe(III)-OH- EPR signal. In contrast, DCMU does not decrease this signal and does not influence its photochemical changes at 140 K. Thus, the EPR spectrum of Cyt b-559 Fe(III) ligated by OH- is not affected by changes at the acceptor side of Photosystem 11, and its photochemical decrease is probably not due to reduction via the acceptor side. Comparison of the effect of KF on the model heme Fe(III) in myoglobin (Mb) at pH 10.5 shows that F- replaces OH- as a ligand at the sixth coordination position of the heme Fe(III) in both Mb and chloroplasts Cyt b-559. This replacement is accompanied by changes of the symmetry of the molecular field causing a disappearance of the EPR signals at g = 6.8 and 5.0. Our results provide further evidence for a possible participation of the Cyt b-559 heme Fe ligated by OH- in photosynthetic water oxidation.
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