An Oxyl/oxo Mechanism for Oxygen-oxygen Coupling in PSII Revealed by an X-ray Free-electron Laser

Photosynthetic water oxidation is catalyzed by the MnCaO cluster of photosystem II (PSII) with linear progression through five S-state intermediates (S to S). To reveal the mechanism of water oxidation, we analyzed structures of PSII in the S, S, and S states by x-ray free-electron laser serial crystallography. No insertion of water was found in S, but flipping of D1 Glu upon transition to S leads to the opening of a water channel and provides a space for incorporation of an additional oxygen ligand, resulting in an open cubane MnCaO cluster with an oxyl/oxo bridge. Structural changes of PSII between the different S states reveal cooperative action of substrate water access, proton release, and dioxygen formation in photosynthetic water oxidation.