Peripheral Light-harvesting Antenna Research Articles

Light Harvesting in Photosystem I Supercomplexes,

In photosynthetic membranes of cyanobacteria, algae, and higher plants, photosystem I (PSI) mediates light-driven transmembrane electron transfer from plastocyanin or cytochrome c6 to the ferredoxin-NADP complex. The oxidoreductase function of PSI is sensitized by a reversible photooxidation of primary electron donor P700, which launches a multistep electron transfer via a series of redox cofactors of the reaction center (RC). The excitation energy for the functioning of the primary electron donor in the RC is delivered via the chlorophyll core antenna in the complex with peripheral light-harvesting antennas. Supermolecular complexes of the PSI acquire remarkably different structural forms of the peripheral light-harvesting antenna complexes, including distinct pigment types and organizational principles. The PSI core antenna, being the main functional unit of the supercomplexes, provides an increased functional connectivity in the chlorophyll antenna network due to dense pigment packing resulting in a fast spread of the excitation among the neighbors. Functional connectivity within the network as well as the spectral overlap of antenna pigments allows equilibration of the excitation energy in the depth of the whole membrane within picoseconds and loss-free delivery of the excitation to primary donor P700 within 20-40 ps. Low-light-adapted cyanobacteria under iron-deficiency conditions extend this capacity via assembly of efficiently energy coupled rings of CP43-like complexes around the PSI trimers. In green algae and higher plants, less efficient energy coupling in the eukaryotic PSI-LHCI supercomplexes is probably a result of the structural adaptation of the Chl a/b binding LHCI peripheral antenna that not only extends the absorption cross section of the PSI core but participates in regulation of excitation flows between the two photosystems as well as in photoprotection.

Different Sensitivities to Oxygen Between Two Strains of the Photosynthetic Green Sulfur Bacterium Chlorobium vibrioforme NCIB 8327 with Bacteriochlorophyll c and d

Two sub-strains of the anoxygenic photosynthetic green sulfur bacterium Chlorobium vibrioforme NCIB 8327 were derived from the same clone and could be discriminated only by their possession of either bacteriochlorophyll (BChl) c or d as the major pigment in the peripheral light-harvesting antenna system, chlorosome (Saga Y et al. (2003) Anal Sci 19: 1575-1579). In the presence of a proper amount of oxygen in the initial culture medium, the BChl d strain showed longer retardation on its growth initiation than the BChl c strain, indicating that the latter was advantageous for survival under aerobic light conditions which produced reactive oxygen species in vivo. The result would be ascribable to the difference of the midpoint potentials between two kinds of chlorosomes formed by self-aggregates of BChl c and d as measured by their fluorescence quenching.

Photo-conversion of chlorophylls in higher-plant CP43 characterized by persistent spectral hole burning at 1.7 K

The chlorophyll–protein complex CP43 is a proximal-antenna subunit important for excitation energy transfer from peripheral light-harvesting antenna to the Photosystem II (PSII) reaction centre. We report persistent spectral hole burning at 1.7 K in the Q y (0,0) origin and Q y (1,0) vibrational satellite bands of chlorophyll a (chl a) in CP43 isolated from higher plants. The isolated CP43 is known to possess two quasi-degenerate ‘red’ trap states. We find persistent hole burning in the primary trap exhibits a photoconversion property, with a photoproduct located well outside its inhomogeneous distribution. This photoconversion of trap chl a molecules also occurs with non-selective white light illumination. The contribution of the ‘red’ states to CP43 absorption is discussed.

Excitonic coupling strength and coherence length in the singlet and triplet excited states of meso-meso directly linked Zn(II)porphyrin arrays.

Excitation-energy transport (EET) phenomena in meso-meso directly linked Zn(II)porphyrin arrays in the singlet and triplet excited states were investigated with a view to electronic coupling strength and coherence length by steady-state and time-resolved spectroscopic measurements. To investigate energy transfer in the triplet states, we modified the Zn(II)porphyrin arrays with bromo substituents at both ends. The coupling strength of the Soret bands of the arrays was estimated to be about 2200 cm-1, and that of the Q bands is about 570 cm-1. The coherence length in the S1 state of the Zn(II)porphyrin arrays was determined to be 4-5 porphyrin units, which is comparable to that of the well-ordered two-dimensional circular structure B850 in the peripheral light-harvesting antenna (LH2) in photosynthetic purple bacteria. This indicates that the Zn(II)porphyrin arrays are well suited for mimicking natural light-harvesting antenna complexes. On the other hand, the rate of energy transfer in the triplet state is estimated to be on the order of 100 microseconds-1, and the very weak coupling between the triplet states (ca. 0.003 cm-1), indicates that the triplet excitation energy is essentially localized on a single porphyrin moiety.

Excitation Energy Transfer in the Lhca1 Subunit of LHC I-730 Peripheral Antenna of Photosystem I

Femtosecond transient absorption spectroscopy at 77 K was used to study excitation energy transfer processes in the reconstituted (r-)Lhca1 subunit of the peripheral light-harvesting antenna complex LHCI-730 of Photosystem I from higher plants. Under selective excitation of chlorophyll (Chl) b at 645 nm with 200 fs laser pulses, two energy transfer processes from Chl b to spectral forms of Chl a with lifetimes of 500 fs and 2.8 ps are induced. The subpicosecond phase reflects the energy transfer between closely located Chl b and Chl a molecules. Excitation at 670 nm induces a similar ultrafast process of energy transfer between neighboring Chl a molecules with a lifetime of 300 fs. The kinetic processes with the lifetime of 3−4 ps indicate an excitation equilibration among pools of Chl b and Chl a in the r-Lhca1, which involves the longest-wavelength spectral form of Chl a at 684 nm. This is in contrast to the Lhca4 subunit of the LHCI-730 heterodimer, which binds the longest-wavelength (red) pigments absorbing at 700−705 nm and responsible for the red fluorescence at 735 nm in LHCI. The spectral pattern of transient absorption changes in the r-Lhca1 is similar to that of the minor light-harvesting complex CP29 of PSII with which the Lhca1 shares significant similarity in protein sequences. A tentative assignment of the observed Chl spectral forms to the pigments in the Lhca1 subunit is presented based on the derivative absorption spectroscopy, transient absorption spectra, and the suggestion that the pigment−protein interactions dominate the spectral heterogeneity of the Qy transitions of Chl a and b.

Molecular origin of red pigments in a peripheral light-harvesting antenna of Photosystem I: ultrafast absorption spectroscopy of recombinant Lhca4

Molecular origin of red pigments in a peripheral light-harvesting antenna of Photosystem I: ultrafast absorption spectroscopy of recombinant Lhca4

Arrangement of photosystem II supercomplexes in crystalline macrodomains within the thylakoid membrane of green plant chloroplasts

The chloroplast thylakoid membrane of green plants is organized in stacked grana membranes and unstacked stroma membranes. We investigated the structural organization of Photosystem II (PSII) in paired grana membrane fragments by transmission electron microscopy. The membrane fragments were obtained by a short treatment of thylakoid membranes with the mild detergent n-dodecyl-α, d-maltoside and are thought to reflect the grana membranes in a native state. The membranes frequently show crystalline macrodomains in which PSII is organized in rows spaced by either 26.3 nm (large-spaced crystals) or 23 nm (small-spaced crystals). The small-spaced crystals are less common but better ordered. Image analysis of the crystals by an aperiodic approach revealed the precise positions of the core parts of PSII in the lattices, as well as features of the peripheral light-harvesting antenna. Together, they indicate that the so-called C 2S 2 and C 2S 2M supercomplexes form the basic motifs of the small-spaced and large-spaced crystals, respectively. An analysis of a pair of membranes with a well-ordered large-spaced crystal reveals that many PSII complexes in one layer face only light-harvesting complexes (LHCII) in the other layer. The implications of this type of organization for the efficient transfer of excitation energy from LHCII to PSII and for the stacking of grana membranes are discussed.

Structural analysis of photosystem II in far-red-light-adapted thylakoid membranes. New crystal forms provide evidence for a dynamic reorganization of light-harvesting antennae subunits.

We studied two-dimensional crystals of the major pigment-protein complex, photosystem II, in far-red-light-adapted thylakoid membranes of the viridis-zb63 mutant of barley. Significantly larger grana membranes were produced with an increased synthesis of the entire photosystem II complex. These red-light-adapted membranes also contained two-dimensional crystals with a high frequency. Three different crystal forms of photosystem II were observed, providing the following data which further our understanding of the architecture of the native complex. (a) The oligomeric form of photosystem II in the membrane was monomeric in all crystal forms, but with a clear non-crystallographic pseudo-twofold symmetry. This was more apparent on the lumenal face of the complex. (b) The variability of unit cell contacts in different crystal forms implied that the peripheral light-harvesting antenna complex and the core of the complex were loosely connected. These peripheral subunits were predicted to rearrange so that they can either encircle the core complex or associate in parallel channels separated by lines of core complexes. (c) Grana membranes were found to retain a double-layered inside-out character, with a stromal face-to-stromal face packing. However, the presence of a crystal in one membrane did not necessarily impose crystallinity on its pair.

Temperature Dependence of Excitation Transfer in LH2 of Rhodobacter sphaeroides

Using two-color pump−probe femtosecond spectroscopy, the temperature dependence of the energy transfer rate within the peripheral light-harvesting antenna (LH2) of the photosynthetic bacterium Rhodobacter sphaeroides has been measured. The energy transfer time from B800 to B850 is determined to be 0.7, 1.2, and 1.5 ps at 300, 77, and 4.2 K, respectively. These data, combined with earlier results, have been analyzed with regard to the crystal structure and spectroscopic properties of the purple bacterial LH2 complex. We conclude that the transfer within B800 occurs mainly via the incoherent Förster hopping mechanism. For B800 to B850 transfer, estimates based on the Förster formula yield considerably slower transfer times than experimentally observed, suggesting that an additional mechanism may be involved in enhancing the transfer rate. We suggest two possibilities: transfer via the upper excitonic component of B850 band and/or transfer mediated by a carotenoid molecule.

One- and two-exciton bands in the LH2 antenna of Rhodopseudomonas acidophila

The properties of one- and two-exciton states of the circular aggregate responsible for the B850 band in the peripheral light harvesting antenna (LH2), extracted from the purple bacterium Rhodopseudomonas acidophila/strain 10050, have been studied at room temperature by non-linear absorption, differential optical density spectroscopy as well as time-resolved fluorescence spectroscopy. As a main result, giant dipole moments for the transition between ground state and the one-exciton band as well as for the transition between the one-exciton band and the two-exciton band have been estimated, corresponding to a delocalization of the excitation energy over most if not all of the 18 BChl a molecules of the aggregate.

Electronic Excitation Transfer in the LH2 Complex of Rhodobacter sphaeroides

Ultrafast fluorescence upconversion measurements were carried out on the peripheral (LH2) light harvesting antenna complex of Rhodobacter sphaeroides isolated in the detergents N-octyl-β-d-glucopyranoside and lauryl dimethylamine oxide. The B800 and B850 bands were excited in separate experiments, and the B850 emission was detected in each case. We make use of the recently determined crystal structure of a purple bacterial LH2 complex to simulate our data and to calculate the exciton level structure of the B850 aggregate. The B800 to B850 excitation transfer occurs with a 650 fs time constant. The depolarization of B850 emission follows a wavelength dependent, biexponential decay with time constants 50−90 and 400−500 fs. We can reproduce the non-exponentiality of the depolarization by assuming incoherent hopping between dimeric sites with a 250 cm-1 full width at half-maximum (fwhm) Gaussian distribution of site energies (inhomogeneity). We calculate the homogeneous hopping time between dimers in B850 to ...

Exciton Delocalization Length in the B850 Antenna of Rhodobacter sphaeroides

The properties of elementary excited states in the B850 band of the peripheral light-harvesting antenna (LH2) of the photosynthetic purple bacterium Rhodobacter sphaeroides has been studied at room temperature by means of femtosecond transient absorption experiments combined with computer simulations. Polarized pump−probe kinetics have a fast component of 100 and 65 fs for the anisotropic and isotropic decays, respectively. Direct numerical simulations show that for incoherent hopping-like excitation transfer in the B850 ring of 18 Bchl a molecules at room temperature the fast component of the anisotropy decay is 3 times longer than the corresponding component of the isotropic decay, strongly suggesting that delocalized exciton states are involved in the observed dynamics. To estimate the coherence length of the exciton we have measured absorption difference spectra of LH2 from 810 to 880 nm 2 ps after the excitation into the B800 band with 75 fs laser pulses. Exciton calculations where also monomeric dou...

Energy transfer in spectrally inhomogeneous light-harvesting pigment-protein complexes of purple bacteria

Energy transfer within the peripheral light-harvesting antenna of the purple bacteria Rhodobacter sphaeroides and Rhodopseudomonas palustris was studied by one- and two-color pump-probe absorption spectroscopy with approximately 100-fs tunable pulses at room temperature and at 77 K. The energy transfer from B800 to B850 occurs with a time constant of 0.7 +/- 0.05 ps at room temperature and 1.8 +/- 0.2 ps at 77 K and is similar in both species. Anisotropy measurements suggest a limited but fast B800 <--> B800 transfer time (tau approximately 0.3 ps). This is analyzed as incoherent hopping of the excitation in a system of spectrally inhomogeneous antenna pigment-protein complexes, by a master equation approach. The simulations show that the measured B800 dynamics is well described as energy transfer with a characteristic average nearest-neighbor pairwise transfer time of 0.35 ps among approximately 10 Bchl molecules in a circular arrangement, in good agreement with the recent high-resolution structure of LH2. The possible presence of fast intramolecular relaxation processes within the Bchl a molecule was investigated by measurement of time-resolved difference absorption spectra and kinetics of Bchl a in solution and in low-temperature glasses. From these measurements it is concluded that fast transients observed at room temperature are due mainly to solvation processes, whereas at 77 K predominantly slower (> 10-ps) relaxation occurs.

Photosynthetic water oxidation: The protein framework

Approximately 20 protein subunits are associated with the PS II complex, not counting subunits of peripheral light-harvesting antenna complexes. However, it is not yet established which proteins specifically are involved in the water-oxidation process. Much evidence supports the concept that the D1/D2 reaction center heterodimer not only plays a central role in the primary photochemistry of Photosystem II, but also is involved in electron donation to P680 and in ligation of the manganese cluster. This evidence includes (a) the primary donor to P680 has been shown to be a redox-active tyrosyl residue (Tyr161) in the D1 protein, and (b) site-directed mutagenesis and computer-assisted modeling of the reaction center heterodimer have suggested several sites with a possible function in manganese ligation. These include Asp170, Gln165 and Gln189 of the D1 protein and Glu69 of the D2 protein as well as the C-terminal portion of the mature D1 protein. Also, hydrophilic loops of the chlorophyll-binding protein CP43 that are exposed at the inner thylakoid surface could be essential for the water-splitting process.In photosynthetic eukaryotes, three lumenal extrinsic proteins, PS II-O (33 kDa), PS II-P (23 kDa) and PS II-Q (16 kDa), influence the properties of the manganese cluster without being involved in the actual catalysis of water oxidation. The extrinsic proteins together may have multiple binding sites to the integral portion of PS II, which could be provided by the D1/D2 heterodimer and CP47. A major role for the PS II-O protein is to stabilize the manganese cluster. Most experimental evidence favors a connection of the PS II-P protein with binding of the Cl(-) and Ca(2+) ions required for the water oxidation, while the PS II-Q protein seems to be associated only with the Cl(-) requirement. The two latter proteins are not present in PS II of prokaryotic organisms, where their functions may be replaced by a 10-12 kDa subunit and a newly discovered low-potential cytochrome c-550.

Chlorophyll-protein derivative of the peripheral light-harvesting antenna of Photosystem I

Chlorophyll-protein derivative of the peripheral light-harvesting antenna of Photosystem I