The crystal structure at 4.8 angstrom resolution of the reaction center-light harvesting 1 (RC-LH1) core complex from Rhodopseudomonas palustris shows the reaction center surrounded by an oval LH1 complex that consists of 15 pairs of transmembrane helical alpha- and beta-apoproteins and their coordinated bacteriochlorophylls. Complete closure of the RC by the LH1 is prevented by a single transmembrane helix, out of register with the array of inner LH1 alpha-apoproteins. This break, located next to the binding site in the reaction center for the secondary electron acceptor ubiquinone (UQB), may provide a portal through which UQB can transfer electrons to cytochrome b/c1.

Photosynthetic organisms flourish under low light intensities by converting photoenergy to chemical energy with near unity quantum efficiency and under high light intensities by safely dissipating excess photoenergy and deleterious photoproducts. The molecular mechanisms balancing these two functions remain incompletely described. One critical barrier to characterizing the mechanisms responsible for these processes is that they occur within proteins whose excited-state properties vary drastically among individual proteins and even within a single protein over time. In ensemble measurements, these excited-state properties appear only as the average value. To overcome this averaging, we investigate the purple bacterial antenna protein light harvesting complex 2 (LH2) from Rhodopseudomonas acidophila at the single-protein level. We use a room-temperature, single-molecule technique, the antiBrownian electrokinetic trap, to study LH2 in a solution-phase (nonperturbative) environment. By performing simultaneous measurements of fluorescence intensity, lifetime, and spectra of single LH2 complexes, we identify three distinct states and observe transitions occurring among them on a timescale of seconds. Our results reveal that LH2 complexes undergo photoactivated switching to a quenched state, likely by a conformational change, and thermally revert to the ground state. This is a previously unobserved, reversible quenching pathway, and is one mechanism through which photosynthetic organisms can adapt to changes in light intensities.photosynthesis | purple bacteria | fluorescence spectroscopy I n photosynthetic light harvesting, networks of antenna pigmentprotein complexes (PPCs) absorb sunlight, then efficiently transport the excitation through these networks to the reaction center, a PPC dedicated to charge separation (1, 2). Photosynthetic systems can complete this process both with near unity quantum efficiency and also function at light levels that provide photoenergy in excess of the capacity of downstream photochemistry. This is accomplished through mechanisms that dissipate harmful by-products produced by unused photoenergy (2, 3). The absorption, energy transport, and dissipation properties are governed by the balance of pigment-pigment and pigment-protein couplings (4). However, the molecular machinery responsible for this balance, and for the couplings themselves, is still poorly understood. This is because the couplings are highly sensitive to intermolecular distances. As a result of this sensitivity, the light-harvesting properties vary drastically among individual PPCs, because of small differences in protein conformation, and vary drastically even within a single PPC over time, because of protein fluctuations (2). In ensemble measurements, these drastic variations appear solely as a static, average value. Therefore, only through single-molecule spectroscopy can we investigate the photodynamics of individual PPCs, specifically how the absorption, energy transport, and dissipation of each change with time. We ...

Energy transfer and trapping in the light harvesting antennae of purple photosynthetic bacteria is an ultrafast process, which occurs with a quantum efficiency close to unity. However the mechanisms behind this process have not yet been fully understood. Recently it was proposed that low-lying energy dark states, such as charge transfer states and polaron pairs, play an important role in the dynamics and directionality of energy transfer. However, it is difficult to directly detect those states because of their small transition dipole moment and overlap with the B850/B870 exciton bands. Here we present a new experimental approach, which combines the selectivity of two-dimensional electronic spectroscopy with the availability of genetically modified light harvesting complexes, to reveal the presence of those dark states in both the genetically modified and the wild-type light harvesting 2 complexes of Rhodopseudomonas palustris. We suggest that Nature has used the unavoidable charge transfer processes that occur when LH pigments are concentrated to enhance and direct the flow of energy.

We report quantum chemical calculations using multireference perturbation theory (MRPT) with the density matrix renormalization group (DMRG) plus photothermal deflection spectroscopy measurements to investigate the manifold of carotenoid excited states and establish their energies relative to the bright state (S 2 ) as a function of nuclear reorganization. We conclude that the primary photophysics and function of carotenoids are determined by interplay of only the bright (S 2 ) and lowest-energy dark (S 1 ) states. The lowest-lying dark state, far from being energetically distinguishable from the lowest-lying bright state along the entire excited-state nuclear reorganization pathway, is instead computed to be either the second or first excited state depending on what equilibrium geometry is considered. This result suggests that, rather than there being a dark intermediate excited state bridging a non-negligible energy gap from the lowestlying dark state to the lowest-lying bright state, there is in fact no appreciable energy gap to bridge following photoexcitation. Instead, excited-state nuclear reorganization constitutes the bridge from S 2 to S 1 , in the sense that these two states attain energetic degeneracy along this pathway.

Evidence for the formation of self-trapped exciton states in photosynthetic antenna complexes is provided by comparing single-molecule fluorescence-excitation and emission spectra that have been recorded from the same individual LH2 complex from Rhodopseudomonas acidophila . While the excitation spectra showed the signatures for the B800 and B850 bands as observed previously, two distinctively different types of emission spectra were found. One group of antenna complexes shows spectra with a relatively narrow spectral profile with a clear signature of a zero-phonon line, whereas the other group of complexes displays spectra that consist only of a broad featureless band. Analysis of these data reveals clear correlations between the spectral position of the emission, the width of the spectral profile, and the associated electron-phonon coupling strength.