1,720,999 research outputs found

    The size of the population of weakly coupled chlorophyll pigments involved in thylakoid photoinhibition determined by steady-state fluorescence spectroscopy

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    AbstractOn the basis of experiments with singlet quenchers and in agreement with previous data, it is suggested that a population of energetically weakly coupled chlorophylls may play a central role in photoinhibition in vivo and in vitro. In the present study, we have used steady state fluorescence techniques to gain direct evidence for these uncoupled chlorophylls. Due to the presence of their emission maxima, near 650 nm and more prominently in the 670–675 nm interval both chlorophylls b and a seem to be involved. A straightforward mathematical model is developed to describe the data which allows us to conclude that the uncoupled/weakly coupled population size is in the range of 1–3 molecules per photosystem

    The Q(y) absorption transition of the light harvesting complex II as determined by structure based analysis of chlorophyll nmacrocycle deforemations

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    The absorption spectrum of the main antenna complex of photosystem II, LHCII, has been modeled using, as starting points, the chlorophyll (chl) atomic coordinates as obtained by the LHCII crystal analysis [Liu, Z., Yan, H., Wang, K., Kuang, T., Zhang, J., Gui, L., An, X., and Chang, W. (2004) Nature 428, 287-292] of three different trimers. The chl site Q(y) transition energies have been obtained in terms of the chl macrocycle deformations influencing the energy level of the chl frontier orbitals. Using these chl site transition energy values and the entire set of interaction energies, calculated in the ideal dipole approximation, the complete Hamiltonians for the three LHCII trimers have been written and the full set of 42 eigenstates of each LHCII trimer have been calculated. With the 42 transition energies and transition dipole strengths, either unperturbed or associated to the eigenstates, the LHCII Q(y) absorption spectrum has been calculated using a chl absorption band shape. These calculations have been performed both in vacuo and in the presence of a medium. Despite the number of approximations, a good correlation with the measured absorption spectrum of a LHCII preparation is obtained. This analysis shows that, although a substantial C3 symmetry of the LHCII trimer in terms of both chl-chl distances and interaction energies is present, a marked variation among monomer subsets of site transition energies is estimated. This leads to a C3 symmetry breaking in the unperturbed chl site transition energies set and, consequently, in the trimer eigenstates. It is also concluded that interactions among chlorophylls do not significantly modify the light absorption role of LHCII in plant leaves

    On phytochrome absorption and the phytochrome photoequilibrium in a green leaf: environmental sensitivity and photoequilibrium time

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    The average, corrected attenuance spectra for both spectral forms of phytochrome in a mature leaf were calculated. Optical masking by chlorophyll together with the detour effect (optical path lengthening effect) due to multiple light scattering led to large changes in both the Qy band shape and wavelength position and the effective intensity of the weak vibrational bands increases. The Pfr/Pr oscillator-strength-ratio between 400-750 nm (0.93 in vitro), becomes 1.63 in a leaf. Thus the dominant absorption form is Pfr. These two values permit calculation of the phytochrome photoequilibrium under conditions of daylight illumination both in vitro and in folia. These values are 0.6 and 0.38 respectively. Previous literature estimates for the situation in vitro, based on the 660/730 nm absorption ratio, yielded values close to 0.6. It is demonstrated that this large decrease in the phytochrome photoequilibrium in a leaf has the effect of translating this parameter to a position on the dose (red/far-red light ratio)-response (Pfr/Ptot) plot towards greater sensitivity to changes in the environmental red/far-red ratio. The increased sensitivity factor is almost five-fold for the daylight environment and is even greater for the various shade-light environments. The approximate time taken to attain photoequilibrium (1/e lifetime) has also been calculated for phytochrome in a leaf in different light environments. For the daylight environment the photoequilibration time is 5 s, which increases into the 20-80 s interval under different degrees of shade light. Thus, despite the strong optical masking by chlorophyll in a mature leaf, the phytochrome photoequilibrium is attained quite rapidly on a physiological time scale

    A study of the relation between CP29 phosphorylation, zeaxanthin content and fluorescence quenching parameters in Zea mays leaves

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    The hypothesis that phosphorylation or the minor photosystem II antenna complex CP29 (CP34 formation) in Zea mays (cv. Dekalb DK300). under conditions of illumination and low temperature stress, may constitute a protective mechanism against photoinhibition. has been investigated. It is demonstrated that illumination at low temperature induces a marked increase in reversible non-photochemical quenching yield of chlorophyll fluorescence, together with CP34 formation, These two parameters, however, are not related as CP34 dephosphorylates to CP29 in the dark, with a half-time of about 10 min, while tho enhanced non-photochemical quenching yield is stable for many hours. The enhanced non-photochemical quenching yield seems to correlate with zeaxanthin formation. The influence of CP34 formation on photoinhibition was also directly investigated. No measurable effect on this parameter could be observed after treatment with high light. It is concluded that CP34 is probably not directly involved in photoprotective processes

    Reconstituted CP29 : multicomponent fluorescence decay from an optically homogeneous sample

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    The multiexponential fluorescence decay of the CP29 complex in which the apoprotein and pigments were reconstituted in vitro was examined. Of the three decay components observed only the two dominant ones, with about 3 and 5 ns lifetimes, were studied. The main question addressed was whether the multicomponent decay was associated with sample optical heterogeneity. To this end, we examined the optical absorption and fluorescence of the CP29 sample by means of two different and independent experimental strategies. This approach was used as the wavelength positions of the absorption/fluorescence spectral forms has recently been shown to be a sensitive indicator of the binding site-induced porphyrin ring deformation (Zucchelli et al. Biophys J 93:2240-2254, 2007) and hence of apoprotein conformational changes. The data indicate that this CP29 sample is optically homogeneous. It is hypothesised that the different lifetimes are explained in terms of multiple detergent/CP29 interactions leading to different quenching states, a suggestion that allows for optical homogeneity

    Fluorescence Lifetime Spectrum of the Plant Photosystem II Core Complex : Photochemistry Does Not Induce Specific Reaction Center Quenching

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    The photosystem II kinetic model (diffusion or trap-limited) is still much debated. There is discussion about whether energy transfer from the core antenna (CP47 and CP43) to the reaction center complex (D1-D2-cyt b559) is rate-limiting (transfer to trap-limited). This study investigates this problem in isolated core particles by exploiting the different optical properties of the core antenna and the reaction center complex near 680 nm, due to P680 and an isoenergetic pheophytin. This was used as a marker feature for the reaction center complex. If the transfer to the trap-limited model were correct, assuming excited-state thermalization, the specific reaction center fluorescence decay lifetime should be shorter near 680 nm, where there is reaction center complex specificity, than at the other emission wavelengths. Such a selective reaction center feature was not observed in fluorescence decay measurements. At the experimental resolution used here, we conclude that the trap-limited energy transfer to the reaction center could, at the most, be 20% limiting. Thus, the transfer to the trap-limited model is not supported. A kinetic, compartmental analysis was also performed on the data, taking into account a large number of separate measurements and the associated errors. Target analysis, considering these intermeasurement errors, yielded two minima which adequately describe the fluorescence lifetime data. The nonunique nature of the description is due to the fact that we have taken into consideration these intermeasurement errors. In our case, due to these errors, a correct kinetic model interpretation required additional experimental information

    Photoinhibition in vivo and in vitro involves weakly coupled chlorophyll-protein complexes

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    In the present study the analysis of the relation between the excited state population in the photosystem 11 (PSII) antenna and photoinactivation has been extended from an in vitro system, isolated thylakoids, to an in vivo system, Chlamydomonas reinhardtii cells. The results indicate that the excited state quenching by an added singlet quencher induces maximal protection against photoinhibition of about 30% of that expected on the basis of the observed light intensity-treatment time reciprocity rule. Similar results, obtained previously with thylakoids, have been interpreted in terms of damaged or incorrectly assembled complexes that play an important role in photoinhibition in the thylakoid membranes (Santabarbara, S., K. Neverov, F. M. Garlaschi, G. Zucchelli and R. C. Jennings [2001] Involvement of uncoupled antenna chlorophylls in photoinhibition in thylakoids. FEBS Lett. 491, 109-113.). In an attempt to better define this aspect, the photoinhibition action spectra were determined for mutant barley thylakoids, lacking the chlorophyll (Chl) a-b complexes of the outer antenna, and for its wild type. The results indicate that in both systems the action spectra are significantly blueshifted (2-4 nm) and are broader than the PSII absorption in the membranes. These data are interpreted in terms of a heterogeneous population of outer and inner antenna pigment-protein complexes that contain significant levels of uncoupled Chl

    Chlorophyll Ring Deformation Modulates Qy Electronic Energy in Chlorophyll-Protein Complexes and Generates Spectral Forms

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    AbstractThe possibility that the chlorophyll (chl) ring distortions observed in the crystal structures of chl-protein complexes are involved in the transition energy modulation, giving rise to the spectral forms, is investigated. The out-of-plane chl-macrocycle distortions are described using an orthonormal set of deformations, defined by the displacements along the six lowest-frequency, out-of-plane normal coordinates. The total chl-ring deformation is the linear combination of these six deformations. The two higher occupied and the two lower unoccupied chl molecular orbitals, which define the Qy electronic transition, have the same symmetry as four of the six out-of-plane lowest frequency modes. We assume that a deformation along the normal-coordinate having the same symmetry as a given molecular orbital will perturb that orbital and modify its energy. The changes in the chl Qy transition energies are evaluated in the Peridinin-Chl-Protein complex and in light harvesting complex II (LHCII), using crystallographic data. The macrocycle deformations induce a distribution of the chl Qy electronic energy transitions which, for LHCII, is broader for chla than for chlb. This provides the physical mechanism to explain the long-held view that the chla spectral forms in LHCII are both more numerous and cover a wider energy range than those of chlb

    Going Beyond Counting First Authors in Author Co-citation Analysis

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    The present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
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