Chloride-depletion effects in the calcium-deficient oxygen-evolving complex of photosystem II

The effects of Cl-depletion in photosystem II (PS-II)-enriched membranes have been investigated by electron paramagnetic resonance (EPR) spectroscopy after removal of the 17- and 23-kDa polypeptides and depletion of Ca2+ by NaCl treatment. When the salt treatment was done in the presence of a high concentration (5 mM) of the chelator [ethylenebis(oxyethylenenitrilo)]tetraacetic acid (EGTA), a modified dark-stable multiline signal was observed from the S2 state and a 13 mT wide S3 signal could be generated by illumination at 0 degrees C as reported previously for experiments conducted under these conditions [Boussac, A., Zimmermann, J.-L., & Rutherford, A. W. (1990) FEBS Lett. 277, 69-74]. The modified S2 multiline signal was lost after a further Cl- depletion in the presence of a low EGTA concentration (50 microM). Upon Cl- reconstitution, a normal S2 multiline signal could be generated by continuous illumination at 200 K. In contrast, a lowering of the EGTA concentration (50 microM) alone, in the presence of Cl- (30 mM), had no effect on the modified S2 multiline signal. These results indicate that the modification of S2 is due to binding of the chelator to PS-II and that Cl- stabilizes the chelator binding. When Cl- depletion in Ca(2+)-depleted PS-II was done in the presence of a high concentration of EGTA (5 mM), the modified S2 multiline signal disappeared but was regenerated by Cl- reconstitution in darkness. These results indicate that when Cl- depletion is done to the EGTA-modified PS-II, the S2 multiline signal disappears but the S2 state remains stable in the dark.(ABSTRACT TRUNCATED AT 250 WORDS)