92 research outputs found

    Modified Gibbs's representation of rotation matrix

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    A modified Gibbs's rotation matrix is derived and the connection with the Euler angles, quaternions, and Cayley−-Klein parameters is established. As particular cases, the Rodrigues and Gibbs parameterizations of the rotation are obtained. The composition law of two rotations from the quaternion representation is presented showing a convenient expression for calculating the successive rotations.Comment: 17 page

    Applying two-photon excitation fluorescence lifetime imaging microscopy to study photosynthesis in plant leaves

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    This study investigates to which extent two-photon excitation (TPE) fluorescence lifetime imaging microscopy can be applied to study picosecond fluorescence kinetics of individual chloroplasts in leaves. Using femtosecond 860 nm excitation pulses, fluorescence lifetimes can be measured in leaves of Arabidopsis thaliana and Alocasia wentii under excitation-annihilation free conditions, both for the F0- and the Fm-state. The corresponding average lifetimes are ~250 ps and ~1.5 ns, respectively, similar to those of isolated chloroplasts. These values appear to be the same for chloroplasts in the top, middle, and bottom layer of the leaves. With the spatial resolution of ~500 nm in the focal (xy) plane and 2 Οm in the z direction, it appears to be impossible to fully resolve the grana stacks and stroma lamellae, but variations in the fluorescence lifetimes, and thus of the composition on a pixel-to-pixel base can be observed

    Linear dichroism and circular dichroism in photosynthesis research

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    The efficiency of photosynthetic light energy conversion depends largely on the molecular architecture of the photosynthetic membranes. Linear- and circular-dichroism (LD and CD) studies have contributed significantly to our knowledge of the molecular organization of pigment systems at different levels of complexity, in pigment–protein complexes, supercomplexes, and their macroassemblies, as well as in entire membranes and membrane systems. Many examples show that LD and CD data are in good agreement with structural data; hence, these spectroscopic tools serve as the basis for linking the structure of photosynthetic pigment–protein complexes to steady-state and time-resolved spectroscopy. They are also indispensable for identifying conformations and interactions in native environments, and for monitoring reorganizations during photosynthetic functions, and are important in characterizing reconstituted and artificially constructed systems. This educational review explains, in simple terms, the basic physical principles, and theory and practice of LD and CD spectroscopies and of some related quantities in the areas of differential polarization spectroscopy and microscopy

    Excitation energy transfer in native and unstacked thylakoid membranes studied by low temperature and ultrafast fluorescence spectroscopy

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    In this work, the transfer of excitation energy was studied in native and cation-depletion induced, unstacked thylakoid membranes of spinach by steady-state and time-resolved fluorescence spectroscopy. Fluorescence emission spectra at 5 K show an increase in photosystem I (PSI) emission upon unstacking, which suggests an increase of its antenna size. Fluorescence excitation measurements at 77 K indicate that the increase of PSI emission upon unstacking is caused both by a direct spillover from the photosystem II (PSII) core antenna and by a functional association of light-harvesting complex II (LHCII) to PSI, which is most likely caused by the formation of LHCII-LHCI-PSI supercomplexes. Time-resolved fluorescence measurements, both at room temperature and at 77 K, reveal differences in the fluorescence decay kinetics of stacked and unstacked membranes. Energy transfer between LHCII and PSI is observed to take place within 25 ps at room temperature and within 38 ps at 77 K, consistent with the formation of LHCII-LHCI-PSI supercomplexes. At the 150-160 ps timescale, both energy transfer from LHCII to PSI as well as spillover from the core antenna of PSII to PSI is shown to occur at 77 K. At room temperature the spillover and energy transfer to PSI is less clear at the 150 ps timescale, because these processes compete with charge separation in the PSII reaction center, which also takes place at a timescale of about 150 ps. Š 2007 Springer Science+Business Media B.V

    Digalactosyl-diacylglycerol-deficiency lowers the thermal stability of thylakoid membranes

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    We investigated the effects of digalactosyl-diacylglycerol (DGDG) on the organization and thermal stability of thylakoid membranes, using wild-type Arabidopsis thaliana and the DGDG-deficient mutant, dgd1. Circular-dichroism measurements reveal that DGDG-deficiency hampers the formation of the chirally organized macrodomains containing the main chlorophyll a/b light-harvesting complexes. The mutation also brings about changes in the overall chlorophyll fluorescence lifetimes, measured in whole leaves as well as in isolated thylakoids. As shown by time-resolved measurements, using the lipophylic fluorescence probe Merocyanine 540 (MC540), the altered lipid composition affects the packing of lipids in the thylakoid membranes but, as revealed by flash-induced electrochromic absorbance changes, the membranes retain their ability for energization. Thermal stability measurements revealed more significant differences. The disassembly of the chiral macrodomains around 55°C, the thermal destabilization of photosystem I complex at 61°C as detected by green gel electrophoresis, as well as the sharp drop in the overall chlorophyll fluorescence lifetime above 45°C (values for the wild type—WT) occur at 4–7°C lower temperatures in dgd1. Similar differences are revealed in the temperature dependence of the lipid packing and the membrane permeability: at elevated temperatures MC540 appears to be extruded from the dgd1 membrane bilayer around 35°C, whereas in WT, it remains lipid-bound up to 45°C and dgd1 and WT membranes become leaky around 35 and 45°C, respectively. It is concluded that DGDG plays important roles in the overall organization of thylakoid membranes especially at elevated temperatures

    The influence of aggregation on triplet formation in light-harvesting chlorophyll a/b pigment-protein complex II of green plants.

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    The influence of aggregation on triplet formation in the light- harvesting pigment-protein complex of photosystem II of green plants (LHCII) has been studied with time-resolved laser flash photolysis. The aggregation state of LHCII has been varied by changing the detergent concentration. The triplet yield increase upon disaggregation and follows the same dependence in the detergent concentration as the flourescence yield. The rate of constant intersystem crossing is not altered by disaggregation, and variations of the triplet yield appear to be due to aggregation-dependent quenching of singlet excited states. The efficiency of triplet transfer in LHCII aggregates from chlorophyll (Chl) to carotenoid (Car) is 92 Âą 7% at room temperature and 82 Âą 6% at 5 K, and does not change upon disaggregation. The Chl's that do not transfer their triplets to Car's seem to be bound to LHCII and are capable of transferring/accepting their singlet excitations to/from other Chl's. Two special contributions of Car triplets are observed: at 525 and 506 nm. Disaggregation of macroaggregates to small aggregates reduces by 10% the relative contribution of Car triplets absorbing at 525 nm. This effect most likely originates from a decreased efficiency of intertrimer Chl-to-Car triplet transfer. At the critical micelle concentration, at which small aggregates are disassembled into trimers, the interactions between Chl and Car are changed. At room temperature, this effect is much more pronounced than at 5 K

    Circular polarized chlorophyll luminescence reflects the macroorganization of grana in pea chloroplasts

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    Circular polarization of luminescence (CPL; Steinberg IZ (1978) Annu Rev Biophys Bioeng 7: 113-137) was applied to study pea chloroplasts in different structural states. The structural changes of chloroplasts were induced by variation of osmotic pressure, concentration of magnesium-ions or photoinhibition. Both large CPL and psi-type circular dichroism (psi, polymerization and salt induced) signals appeared in the presence of granal macrostructure and were sensitive to structural changes of the grana. The relation was studied between the amount of CPL expressed as an emission anisotropy factor
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