pK_a of the ligand water molecules in the oxygen-evolving Mn₄CaO₅ cluster in photosystem II

Release of the protons from the substrate water molecules is prerequisite for O₂ evolution in photosystem II (PSII). Proton-releasing water molecules with low pK_a values at the catalytic moiety can be the substrate water molecules. In some studies, one of the ligand water molecules, W2, is regarded as OH^-. However, the PSII crystal structure shows neither proton acceptor nor proton-transfer pathway for W2, which is not consistent with the assumption of W2 = OH^-. Here we report the pK_a values of the four ligand water molecules, W1 and W2 at Mn4 and W3 and W4 at Ca²⁺, of the Mn₄CaO₅ cluster. pK_a(W1) ≈ pK_a(W2) << pK_a(W3) ≈ pK_a(W4) in the Mn₄CaO₅ cluster in water. However, pK_a(W1) ≈ pK_a(D1-Asp61) << pK_a(W2) in the PSII protein environment. These results suggest that in PSII, deprotonation of W2 is energetically disfavored as far as W1 exists.