New insights into the photochemistry of carotenoid spheroidenone in light-harvesting complex 2 from the purple bacterium Rhodobacter sphaeroides

Light-harvesting complex 2 (LH2) from the semi-aerobically grown purple phototrophic bacterium Rhodobacter sphaeroides was studied using optical (static and time-resolved) and resonance Raman spectroscopies. This antenna complex comprises bacteriochlorophyll (BChl) a and the carotenoid spheroidenone, a ketolated derivative of spheroidene. The results indicate that the spheroidenone-LH2 complex contains two spectral forms of the carotenoid: (1) a minor, ‘‘blue’’ form with an S2 (11 Bu ?) spectral origin band at 522 nm, shifted from the position in organic media simply by the high polarizability of the binding site, and (2) the major, ‘‘red’’ form with the origin band at 562 nm that is associated with a pool of pigments that more strongly interact with protein residues, most likely via hydrogen bonding. Application of targeted modeling of excited-state decay pathways after carotenoid excitation suggests that the high (92%) carotenoid-to-BChl energy transfer efficiency in this LH2 system, relative to LH2 complexes binding carotenoids with comparable double-bond conjugation lengths, derives mainly from resonance energy transfer from spheroidenone S2 (11 Bu ?) state to BChl a via the Qx state of the latter, accounting for 60% of the total transfer. The elevated S2 (11 Bu ?) ? Qx transfer efficiency is apparently associated with substantially decreased energy gap (increased spectral overlap) between the virtual S2 (11 Bu ?) ? S0 (11 Ag -) carotenoid emission and Qx absorption of BChl a. This reduced energetic gap is the ultimate consequence of strong carotenoid–protein interactions, including the inferred hydrogen bondin

Participation of Glutamate-333 of the D1 Polypeptide in the Ligation of the Mn 4 CaO 5 Cluster in Photosystem II

In the 1.9 Å structural model of photosystem II (PDB: 3ARC), the amino acid residue Glu333 of the D1 polypeptide coordinates to the oxygen-evolving Mn4CaO5 cluster. This residue appears to be highly significant in that it bridges the two Mn ions (MnB3 a

The D1-D61N Mutation in <i>Synechocystis</i> sp. PCC 6803 Allows the Observation of pH-Sensitive Intermediates in the Formation and Release of O₂ from Photosystem II

The active site of photosynthetic water oxidation by Photosystem II (PSII) is a manganese–calcium cluster (Mn₄CaO₅). A postulated catalytic base is assumed to be crucial. CP43-Arg357, which is a candidate for the identity of this base, is a second-sphere ligand of the Mn₄–Ca cluster and is located near a putative proton exit pathway, which begins with residue D1-D61. Transient absorption spectroscopy and time-resolved O₂ polarography reveal that in the D1-D61N mutant, the transfer of an electron from the Mn₄CaO₅ cluster to Y_Z^OX and O₂ release during the final step of the catalytic cycle, the S₃–S₀ transition, proceed simultaneously but are more dramatically decelerated than previously thought (<i>t</i>_1/2 of up to ∼50 ms vs a <i>t</i>_1/2 of 1.5 ms in the wild type). Using a bare platinum electrode to record the flash-dependent yields of O₂ from mutant and wild-type PSII has allowed the observation of the kinetics of release of O₂ from extracted thylakoid membranes at various pH values and in the presence of deuterated water. In the mutant, it was possible to resolve a clear lag phase prior to the appearance of O₂, indicating formation of an intermediate before the onset of O₂ formation. The lag phase and the photochemical miss factor were more sensitive to isotope substitution in the mutant, indicating that proton efflux in the mutant proceeds via an alternative pathway. The results are discussed in comparison with earlier results obtained from the substitution of CP43-Arg357 with lysine and in regard to hypotheses concerning the nature of the final steps in photosynthetic water oxidation. These considerations led to the conclusion that proton expulsion during the initial phase of the S₃–S₀ transition starts with the deprotonation of the primary catalytic base, probably CP43-Arg357, followed by efficient proton egress involving the carboxyl group of D1-D61 in a process that constitutes the lag phase immediately prior to O₂ formation chemistry