Fig. 3 a The Mn K-edge spectra of spinach PS II (BBY), from the S0 through S3 states (top) and their second derivative spectra (bottom). The magnitude of the inflection point energy shift for the S0 to S1 (2.1 eV) and S1 and S2 (1.1 eV) is much larger than the shift for the S2 to S3 transition (0.3 eV). The inset shows the pre-edge (1s to 3d transition) from the S-states is enlarged and shown above the Mn K-edge spectra.
b The Fourier transform (FT) from a PS II sample in the S1 state. The three FT Peak I corresponds to Mn-bridging and terminal ligand (N/O) distances at 1.8–2.0 Å, Peak II is from Mn–Mn distances (2 at ~2.7 and 1 at ~2.8 Å), and FT Peak III is from Mn–Mn distance at ~3.3 Å and Mn–Ca distances learn more at ~3.4 Å The EXAFS is interpretable as shells at 1.8 and 2.0 Å (Peak I) attributable to N or O atoms and a shell at ~2.7–2.8 Å (Peak II) from Mn to Mn interactions. An additional shell from Mn was seen at 3.3 Å (Peak III; Fig. 3).
The Mn EXAFS spectra changes upon the S-state transitions, particularly from the S2 to S3 state transition, suggesting that the OEC goes through structural changes triggered by the oxidation state changes and protonation/deprotonation events. Co-factor XAS The S-state catalytic cycle can be studied also by co-factor XAS studies (Cinco et al. 2002). One Ca is known Barasertib to be a part of the OEC, and this has been proven by Ca XAS studies and from X-ray crystallography using Morin Hydrate the anomalous diffraction technique. Regarding Cl, there is no spectroscopic evidence at least in the S1 state that the Cl is a direct ligand to the OEC, although several biochemical studies suggest a critical role for one tightly bound Cl in maintaining oxygen-evolving activity. In general, the requirements of X-ray spectroscopy place some restrictions with respect to sample preparation and experimental
conditions. Ca and Cl in some sense fall into this category. The investigation of light elements can present difficulties due to the presence of an aqueous medium and the pervasive occurrence of C, N, and O in biological materials. In X-ray energy regions, where atmospheric gases absorb, samples must be placed in an atmosphere of helium or in vacuum. For elements like Ca and Cl, which can occur in a wide variety of environments in biological materials, it is particularly challenging to remove sources of background signals that greatly complicate interpreting the results. Another strategy to study the role of such light element co-factor(s) is to replace it with MM-102 heavier element(s). Ca can be replaced chemically or biosynthetically with Sr without losing its enzymatic activity. Similarly, Cl can be substituted with Br. XAS measurements at the Sr K-edge (16,200 eV; Cinco et al. 1998; Pushkar et al. 2008) or Br K-edge (13,600 eV; Haumann et al.