Sergei Savikhin

Singlet Exciton Fission in Polycrystalline Thin Films of a Slip-Stacked Perylenediimide

The crystal structure of N,N-bis(n-octyl)-2,5,8,11-tetraphenylperylene-3,4:9,10-bis(dicarboximide), 1, obtained by X-ray diffraction reveals that 1 has a nearly planar perylene core and π-π stacks at a 3.5 Å interplanar distance in well-separated slip-stacked columns. Theory predicts that slip-stacked, π-π-stacked structures should enhance interchromophore electronic coupling and thus favor singlet exciton fission. Photoexcitation of vapor-deposited polycrystalline 188 nm thick films of 1 results in a 140 ± 20% yield of triplet excitons ((3*)1) in τ(SF) = 180 ± 10 ps. These results illustrate a design strategy for producing perylenediimide and related rylene derivatives that have the optimized interchromophore electronic interactions which promote high-yield singlet exciton fission for potentially enhancing organic solar cell performance and charge separation in systems for artificial photosynthesis.

Electrochromic Shift of Chlorophyll Absorption in Photosystem I from Synechocystis sp. PCC 6803: A Probe of Optical and Dielectric Properties around the Secondary Electron Acceptor

Nanosecond absorption dynamics at approximately 685 nm after excitation of photosystem I (PS I) from Synechocystis sp. PCC 6803 is consistent with electrochromic shift of absorption bands of the Chl a pigments in the vicinity of the secondary electron acceptor A(1). Based on experimental optical data and structure-based simulations, the effective local dielectric constant has been estimated to be between 3 and 20, which suggests that electron transfer in PS I is accompanied by considerable protein relaxation. Similar effective dielectric constant values have been previously observed for the bacterial photosynthetic reaction center and indicate that protein reorganization leading to effective charge screening may be a necessary structural property of proteins that facilitate the charge transfer function. The data presented here also argue against attributing redmost absorption in PS I to closely spaced antenna chlorophylls (Chls) A38 and A39, and suggest that optical transitions of these Chls, along with that of connecting chlorophyll (A40) lie in the range 680-695 nm.

Ultrafast Optical Spectroscopy of Photosystem I

Summary This review discusses energy and electron transfer in Photosystem I (PS I) complexes by means of ultrafast timeresolved optical techniques. In particular, this article addresses directly observable initial (sub)picosecond electronic excitation equilibration among different antenna chlorophyll forms and energy trapping by the charge separation process followed by picosecond electron transfer to the secondary electron acceptor A1. There is still no general agreement on the energy trapping classification in PS I; the validity of diffusion-limited, trap-limited, and mixedenergy trapping models is tested against the available experimental data. Ultrafast experiments on branch-specific complementary mutants of the reaction center open the unique possibility of differentiating between the two highly symmetrical branches of the reaction center, revealing the directionality of electron transfer in PS I. Finally, the most recent optical data questions the conventional sequence of electron transfer steps, and suggests the intriguing possibility that one additional intermediate radical pair may exist that was not previously observed.

On the structural role of the aromatic residue environment of the chlorophyll a in the cytochrome b6f complex

Because light is not required for catalytic turnover of the cytochrome b 6 f complex, the role of the single chlorophyll a in the structure and function of the complex is enigmatic. Photodamage from this pigment is minimized by its short singlet excited-state lifetime ( approximately 200 ps), which has been attributed to quenching by nearby aromatic residues ( Dashdorj et al., 2005). The crystal structure of the complex shows that the fifth ligand of the chlorophyll a contains two water molecules. On the basis of this structure, the properties of the bound chlorophyll and the complex were studied in the cyanobacterium, Synechococcus sp. PCC 7002, through site-directed mutagenesis of aromatic amino acids in the binding niche of the chlorophyll. The b 6 f complex was purified from three mutant strains, a double mutant Phe133Leu/Phe135Leu in subunit IV and single mutants Tyr112Phe and Trp125Leu in the cytochrome b 6 subunit. The purified b 6 f complex from Tyr112Phe or Phe133Leu/Phe135Leu mutants was characterized by (i) a loss of bound Chl and b heme, (ii) a shift in the absorbance peak and increase in bandwidth, (iii) multiple lifetime components, including one of 1.35 ns, and (iv) relatively small time-resolved absorbance anisotropy values of the Chl Q y band. A change in these properties was minimal in the Trp125Leu mutant. In vivo, no decrease in electron-transport efficiency was detected in any of the mutants. It was concluded that (a) perturbation of its aromatic residue niche influences the stability of the Chl a and one or both b hemes in the monomer of the b 6 f complex, and (b) Phe residues (Phe133/Phe135) of subunit IV are important in maintaining the short lifetime of the Chl a singlet excited state, thereby decreasing the probability of singlet oxygen formation.

Spectral Resolution of the Primary Electron Acceptor A0 in Photosystem I

The reduced state of the primary electron acceptor of Photosystem I, A(0), was resolved spectroscopically in its lowest energy Q(y) region for the first time without the addition of chemical reducing agents and without extensive data manipulation. To carry this out, we used the menB mutant of Synechocystis sp. PCC 6803 in which phylloquinone is replaced by plastoquinone-9 in the A(1) sites of Photosystem I. The presence of plastoquinone-9 slows electron transfer from A(0) to A(1), leading to a long-lived A(0)(-) state. This allows its spectral signature to be readily detected in a time-resolved optical pump-probe experiment. The maximum bleaching (A(0)(-) - A(0)) was found to occur at 684 nm with a corresponding extinction coefficient of 43 mM(-1) cm(-1). The data show evidence for an electrochromic shift of an accessory chlorophyll pigment, suggesting that the latter Q(y) absorption band is centered around 682 nm.

Oxygen Concentration Inside a Functioning Photosynthetic Cell

The excess oxygen concentration in the photosynthetic membranes of functioning oxygenic photosynthetic cells was estimated using classical diffusion theory combined with experimental data on oxygen production rates of cyanobacterial cells. The excess oxygen concentration within the plesiomorphic cyanobacterium Gloeobactor violaceus is only 0.025 μM, or four orders of magnitude lower than the oxygen concentration in air-saturated water. Such a low concentration suggests that the first oxygenic photosynthetic bacteria in solitary form could have evolved ∼2.8 billion years ago without special mechanisms to protect them against reactive oxygen species. These mechanisms instead could have been developed during the following ∼500 million years while the oxygen level in the Earths atmosphere was slowly rising. Excess oxygen concentrations within individual cells of the apomorphic cyanobacteria Synechocystis and Synechococcus are 0.064 and 0.25 μM, respectively. These numbers suggest that intramembrane and intracellular proteins in isolated oxygenic photosynthetic cells are not subjected to excessively high oxygen levels. The situation is different for closely packed colonies of photosynthetic cells. Calculations show that the excess concentration within colonies that are ∼40 μm or larger in diameter can be comparable to the oxygen concentration in air-saturated water, suggesting that species forming colonies require protection against reactive oxygen species even in the absence of oxygen in the surrounding atmosphere.

Probing the excitonic landscape of the Chlorobaculum tepidum Fenna-Matthews-Olson (FMO) complex: a mutagenesis approach

In this paper we report the steady-state optical properties of a series of site-directed mutants in the Fenna-Matthews-Olson (FMO) complex of Chlorobaculum tepidum, a photosynthetic green sulfur bacterium. The FMO antenna complex has historically been used as a model system for energy transfer due to the water-soluble nature of the protein, its stability at room temperature, as well as the availability of high-resolution structural data. Eight FMO mutants were constructed with changes in the environment of each of the bacteriochlorophyll a pigments found within each monomer of the homotrimeric FMO complex. Our results reveal multiple changes in low temperature absorption, as well as room temperature CD in each mutant compared to the wild-type FMO complex. These datasets were subsequently used to model the site energies of each pigment in the FMO complex by employing three different Hamiltonians from the literature. This enabled a basic approximation of the site energy shifts imparted on each pigment by the changed amino acid residue. These simulations suggest that, while the three Hamiltonians used in this work provide good fits to the wild-type FMO absorption spectrum, further efforts are required to obtain good fits to the mutant minus wild-type absorption difference spectra. This demonstrates that the use of FMO mutants can be a valuable tool to refine and iterate the current models of energy transfer in this system.

Young research investigators honored at the 2011 Gordon research conference on photosynthesis: ambiance and a perspective

Using photographs taken at the conference site, we provide a perspective on (i) the awards that were given to four young investigators at the 2011 Gordon Research Conference on Photosynthesis, and (ii) the ambiance at this conference, held at Davidson College, North Carolina, during June 12–17, 2011.

Chapter 20:Structure–Function of the Cytochrome b6f Complex: A Design that has Worked for Three Billion Years

The 3.0–3.1 A X-ray structures of the cytochrome b6 f complex from the thermophilic cyanobacterium Mastigocladus laminosus and from the green alga Chlamydomonas reinhardtii are very similar. Eight natural prosthetic groups, four hemes, one [2Fe-2S] cluster, one Chl, one β-carotene, and one n-side plastoquinone are embedded in the eight polypeptide subunits of the complex, four large (18–33 kDa) and four small (∼4 kDa). The complex is organized as a dimer with a molecular weight of 217 kDa in M. laminosus. Other subunits such as ferredoxin: NADP+ reductase may bind transiently and more weakly to the n-side of the complex. Major features of the structure are: (i) a large inter-monomer lipophilic “quinone exchange cavity” that exchanges plastoquinone/quinol with the quinone pool in the lipid bilayer membrane; (ii) a labyrinthine pathway of plastoquinone movement between n- and p-electron exchange sites through the 11 × 12 A portal at the roof of the cavity; (iii) three prosthetic groups with unknown function, a novel high-spin heme (cn) close to heme bn, a chlorophyll a, and a β-carotene; (iv) a proposed function of heme cn is in PS I-linked cyclic electron transport, although the presumed binding site of a “sometime” inhibitor of cyclic ET, antimycin A, is occluded by heme cn; (v) the single Chl a molecule in the monomer is characterized by a short (200 ps) fluorescence lifetime and large anisotropy of fluorescence; and (vi) transfer of energy from the Chl triplet state to the β-carotene occurs despite the 14 A separation of the pigments – it is proposed that this transfer operates through an intraprotein, interpigment O2 channel.

Pathways of Transmembrane Electron Transfer in Cytochrome bc Complexes: Dielectric Heterogeneity and Interheme Coulombic Interactions

The intramembrane cytochrome bc1 complex of the photosynthetic bacterium Rhodobacter capsulatus and the cytochrome b6f complex, which functions in oxygenic photosynthesis, utilize two pairs of b-hemes in a symmetric dimer to accomplish proton-coupled electron transfer. The transmembrane electron transfer pathway in each complex was identified through the novel use of heme Soret band excitonic circular dichroism (CD) spectra, for which the responsible heme-heme interactions were determined from crystal structures. Kinetics of heme reduction and CD amplitude change were measured simultaneously. For bc1, in which the redox potentials of the transmembrane heme pair are separated by 160 mV, heme reduction occurs preferentially to the higher-potential intermonomer heme pair on the electronegative (n) side of the complex. This contrasts with the b6f complex, where the redox potential difference between transmembrane intramonomer p- and n-side hemes is substantially smaller and the n-p pair is preferentially reduced. Limits on the dielectric constant between intramonomer hemes were calculated from the interheme distance and the redox potential difference, ΔEm. The difference in preferred reduction pathway is a consequence of the larger ΔEm between n- and p-side hemes in bc1, which favors the reduction of n-side hemes and cannot be offset by decreased repulsive Coulombic interactions between intramonomer hemes.