Structure and organization of phycobilisomes on membranes of the red alga Porphyridium cruentum

In the present work, electron microscopy and single particle averaging was performed to investigate the supramolecular architecture of hemiellipsoidal phycobilisomes from the unicellular red alga Porphyridium cruentum. The dimensions were measured as 60 × 41 × 34 nm (length × width × height) for randomly ordered phycobilisomes, seen under high-light conditions. The hemiellipsoidal phycobilisomes were found to have a relatively flexible conformation. In closely packed semi-crystalline arrays, observed under low-light conditions, the width is reduced to 31 or 35 nm, about twice the width of the phycobilisome of the cyanobacterium Synechocystis sp. PCC 6803. Since the latter size matches the width of dimeric PSII, we suggest that one PBS lines up with one PSII dimer in cyanobacteria. In red algae, a similar 1:1 ratio under low-light conditions may indicate that the red algal phycobilisome is enlarged by a membrane-bound peripheral antenna which is absent in cyanobacteria

Demonstration of asymmetric electron conduction in pseudosymmetrical photosynthetic reaction centre proteins in an electrical circuit

Photosynthetic reaction centres show promise for biomolecular electronics as nanoscale solar-powered batteries and molecular diodes that are amenable to atomic-level re-engineering. In this work the mechanism of electron conduction across the highly tractable Rhodobacter sphaeroides reaction centre is characterized by conductive atomic force micro-scopy. We find, using engineered proteins of known structure, that only one of the two cofactor wires connecting the positive and negative termini of this reaction centre is capable of conducting unidirectional current under a suitably oriented bias, irrespective of the magnitude of the bias or the applied force at the tunnelling junction. This behaviour, strong functional asymmetry in a largely symmetrical protein–cofactor matrix, recapitulates the strong functional asymmetry characteristic of natural photochemical charge separation, but it is surprising given that the stimulus for electron flow is simply an externally applied bias. Reasons for the electrical resistance displayed by the so-called B-wire of cofactors are explored

Energy migration in Rhodobacter sphaeroides mutants altered by mutagenesis of the peripheral LH2 complex or by removal of the core LH1 complex

AbstractThe photosynthetic apparatus of the purple bacterium Rhodobacter sphaeroides is organised so that light energy absorbed by the peripheral antenna (LH2) complexes migrates towards the core (LH1) complex, before being trapped by the reaction centre (RC). This migration and trapping process has been studied in mutants where the energy levels of the LH2 BChls have been raised by mutagenesis of the C-terminal aromatic residues (Fowler, G.J.S., Visschers, R.W., Grief, G.G., Van Grondelle, R. and Hunter, C.N. (1992) Nature 355, 848–850), and in a mutant which lacks the core complex. In the former case, the alterations to the LH2 complexes did not prevent efficient energy transfer to the LHI-RC complex, but fluorescence emission spectra indicated that the equilibrium of energy within the system was affected so that back transfer from the LH1-RC core is minimised. This mimics the situation found in some other bacteria such as Rhodopseudomonas acidophila and Rps. cryptolactis. In the mutant lacking LH1, energy is transferred from LH2 directly to the RC, despite the absence of the core antenna. Energy transfer efficiencies for carotenoids and LH2 to LH1 were measured for the blue-shifted LH2 mutants, and were found to be high (70%) in each case. These data, together with measurements of excitation annihilation as a function of incident excitation energy, were used to estimate the domain sizes for energy transfer in these mutants. In the LH2 mutants, domains of about 50 to 170 core BChls were found, depending on the type of mutation. One effect of the removal of LH1 appears to be the reorganisation of the peripheral LH2 antenna to form domains of at least 250 BChls

Light-Induced Energetic Decoupling as a Mechanism for Phycobilisome-Related Energy Dissipation in Red Algae: A Single Molecule Study

BACKGROUND: Photosynthetic organisms have developed multiple protective mechanisms to prevent photodamage in vivo under high-light conditions. Cyanobacteria and red algae use phycobilisomes (PBsomes) as their major light-harvesting antennae complexes. The orange carotenoid protein in some cyanobacteria has been demonstrated to play roles in the photoprotective mechanism. The PBsome-itself-related energy dissipation mechanism is still unclear. METHODOLOGY/PRINCIPAL FINDINGS: Here, single-molecule spectroscopy is applied for the first time on the PBsomes of red alga Porphyridium cruentum, to detect the fluorescence emissions of phycoerythrins (PE) and PBsome core complex simultaneously, and the real-time detection could greatly characterize the fluorescence dynamics of individual PBsomes in response to intense light. CONCLUSIONS/SIGNIFICANCE: Our data revealed that strong green-light can induce the fluorescence decrease of PBsome, as well as the fluorescence increase of PE at the first stage of photobleaching. It strongly indicated an energetic decoupling occurring between PE and its neighbor. The fluorescence of PE was subsequently observed to be decreased, showing that PE was photobleached when energy transfer in the PBsomes was disrupted. In contrast, the energetic decoupling was not observed in either the PBsomes fixed with glutaraldehyde, or the mutant PBsomes lacking B-PE and remaining b-PE. It was concluded that the energetic decoupling of the PBsomes occurs at the specific association between B-PE and b-PE within the PBsome rod. Assuming that the same process occurs also at the much lower physiological light intensities, such a decoupling process is proposed to be a strategy corresponding to PBsomes to prevent photodamage of the photosynthetic reaction centers. Finally, a novel photoprotective role of gamma-subunit-containing PE in red algae was discussed

FRAP Analysis on Red Alga Reveals the Fluorescence Recovery Is Ascribed to Intrinsic Photoprocesses of Phycobilisomes than Large-Scale Diffusion

BACKGROUND: Phycobilisomes (PBsomes) are the extrinsic antenna complexes upon the photosynthetic membranes in red algae and most cyanobacteria. The PBsomes in the cyanobacteria has been proposed to present high lateral mobility on the thylakoid membrane surface. In contrast, direct measurement of PBsome motility in red algae has been lacking so far. METHODOLOGY/PRINCIPAL FINDINGS: In this work, we investigated the dynamics of PBsomes in the unicellular red alga Porphyridium cruentum in vivo and in vitro, using fluorescence recovery after photobleaching (FRAP). We found that part of the fluorescence recovery could be detected in both partially- and wholly-bleached wild-type and mutant F11 (UTEX 637) cells. Such partial fluorescence recovery was also observed in glutaraldehyde-treated and betaine-treated cells in which PBsome diffusion should be restricted by cross-linking effect, as well as in isolated PBsomes immobilized on the glass slide. CONCLUSIONS/SIGNIFICANCE: On the basis of our previous structural results showing the PBsome crowding on the native photosynthetic membrane as well as the present FRAP data, we concluded that the fluorescence recovery observed during FRAP experiment in red algae is mainly ascribed to the intrinsic photoprocesses of the bleached PBsomes in situ, rather than the rapid diffusion of PBsomes on thylakoid membranes in vivo. Furthermore, direct observations of the fluorescence dynamics of phycoerythrins using FRAP demonstrated the energetic decoupling of phycoerythrins in PBsomes against strong excitation light in vivo, which is proposed as a photoprotective mechanism in red algae attributed by the PBsomes in response to excess light energy

Transient Absorption Changes of a Degenerate Dimer Induced by Exciton Scattering; Implications for Photosystem II of Photosynthesis

The effect of exciton scattering on transient absorption spectra of a degenerate dimer has been calculated from a simple four-level model. The absorbance changes due to this scattering process not only depend on the polarization of the probe beam, but also on the selectivity of excitation of specific exciton components. In the case of selective excitation, the absorbance changes induced by exciton scattering may play an important role in time-resolved spectroscopy of the reaction center of Photosystem II at short times after excitation