Probing the role of chloride in Photosystem II from Thermosynechococcus elongatus by exchanging chloride for iodide

The active site for water oxidation in Photosystem II (PSII) goes through five sequential oxidation states (S0 to S4) before O2 is evolved. It consists of a Mn4CaO5 cluster and TyrZ, a redox-active tyrosine residue. Chloride ions have been known for long time to be required for the function of the enzyme. However, X-ray data have shown that they are located about 7 Å away from the Mn4CaO5 cluster, a distance that seems too large to be compatible with a direct involvement of chloride in the water splitting chemistry. We have investigated the role of this anion by substituting I− for Cl− in the cyanobacterium Thermosynechococcus elongatus with either Ca2 + or Sr2 + biosynthetically assembled into the Mn4 cluster. The electron transfer steps affected by the exchanges were investigated by time-resolved UV–visible absorption spectroscopy, time-resolved EPR at room temperature and low temperature cw-EPR spectroscopy. In both Ca-PSII and Sr-PSII, the Cl−/I− exchange considerably slowed down the two S3TyrZ
•  → (S3TyrZ
• )′ → S0 reactions in which the fast phase, S3TyrZ
•  → (S3TyrZ
• )′, reflects the electrostatically triggered expulsion of one proton from the catalytic center caused by the positive charge near/on TyrZ
• and the slow phase corresponds to the S0 and O2 formations and to a second proton release. The t1/2 for S0 formation increased from 1.1 ms in Ca/Cl-PSII to ≈ 6 ms in Ca/I-PSII and from 4.8 ms in Sr/Cl-PSII to ≈ 45 ms in Sr/I-PSII. In all cases the TyrZ
• reduction was the limiting step. The kinetic effects are interpreted by a model in which the Ca2 + binding site and the Cl− binding site, although spatially distant, interact. This interaction is likely mediated by the H-bond and/or water molecules network(s) connecting the Cl− and Ca2 + binding sites by which proton release may be channelled.

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► Chloride is substituted for iodide in Photosystem II from T. elongatus.
► Ca/I-PSII and Sr/I-PSII have a high O2 evolving activity.
► The t1/2 of S3TyrZ
• ′ to S0 is 1.1 ms in Ca/Cl-PSII and 6 ms in Ca/I-PSII.
► The t1/2 of S3TyrZ
• ′ to S0 is 4.8 ms in Sr/Cl-PSII and 45 ms in Sr/I-PSII.
► Ca2+ and Cl– interact via the H-bond and/or H2O network(s) connecting the 2 sites.