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Instruments / Technical Area	Publication Details	Published	Actions
B23-Circular Dichroism	Photobleaching of chlorophyll in light-harvesting complex II increases in lipid environment Monika Lingvay , Parveen Akhtar , Krisztina Sebők-Nagy , Tibor Páli , Petar H. Lambrev Frontiers In Plant Science 11 DOI: 10.3389/fpls.2020.00849 Diamond Proposal Number(s): [17698] Open Access Show Abstract ▼ Abstract: Excess light causes damage to the photosynthetic apparatus of plants and algae primarily via reactive oxygen species. Singlet oxygen can be formed by interaction of chlorophyll (Chl) triplet states, especially in the Photosystem II reaction center, with oxygen. Whether Chls in the light-harvesting antenna complexes play direct role in oxidative photodamage is less clear. In this work, light-induced photobleaching of Chls in the major trimeric light-harvesting complex II (LHCII) is investigated in different molecular environments – protein aggregates, embedded in detergent micelles or in reconstituted membranes (proteoliposomes). The effects of intense light treatment were analyzed by absorption and circular dichroism spectroscopy, steady-state and time-resolved fluorescence and EPR spectroscopy. The rate and quantum yield of photobleaching was estimated from the light-induced Chl absorption changes. Photobleaching occurred mainly in Chl a and was accompanied by strong fluorescence quenching of the remaining unbleached Chls. The rate of photobleaching increased by 140% when LHCII was embedded in lipid membranes, compared to detergent-solubilized LHCII. Removing oxygen from the medium or adding antioxidants largely suppressed the bleaching, confirming its oxidative mechanism. Singlet oxygen formation was monitored by EPR spectroscopy using spin traps and spin labels to detect singlet oxygen directly and indirectly, respectively. The quantum yield of Chl a photobleaching in membranes and detergent was found to be 3.4 × 10–5 and 1.4 × 10–5, respectively. These values compare well with the yields of ROS production estimated from spin-trap EPR spectroscopy (around 4 × 10–5 and 2 × 10–5). A kinetic model is proposed, quantifying the generation of Chl and carotenoid triplet states and singlet oxygen. The high quantum yield of photobleaching, especially in the lipid membrane, suggest that direct photodamage of the antenna occurs with rates relevant to photoinhibition in vivo. The results represent further evidence that the molecular environment of LHCII has profound impact on its functional characteristics, including, among others, the susceptibility to photodamage.	Jun 2020
B23-Circular Dichroism	Anisotropic circular dichroism of light-harvesting complex II in oriented lipid bilayers: Theory meets experiment Parveen Akhtar , Dominik Lindorfer , Monika Lingvay , Krzysztof Pawlak , Otto Zsiros , Giuliano Siligardi , Tamas Javorfi , Márta Dorogi , Bettina Ughy , Gyozo Garab , Thomas Renger , Petar H. Lambrev The Journal Of Physical Chemistry B DOI: 10.1021/acs.jpcb.8b12474 Diamond Proposal Number(s): [15094, 16232, 17698, 19120, 20644] Show Abstract ▼ Abstract: Anisotropic circular dichroism (ACD) spectroscopy of macroscopically aligned molecules reveals additional information about their excited states that is lost in the CD of randomly-oriented solutions. ACD spectra of light-harvesting complex II (LHCII)—the main peripheral antenna of photosystem II in plants—in oriented lipid bilayers, were recorded from the far-UV to the visible wavelength region. The ACD spectra show a drastically enhanced magnitude and level of detail compared to the isotropic CD spectra, resolving a greater number of bands and weak optical transitions. Exciton calculations show that the spectral features in the chlorophyll Qy region are well reproduced by an existing Hamiltonian for LHCII, providing further evidence for the identity of energy sinks at chlorophylls a603 and a610 in the stromal layer and chlorophylls a604 and a613 in the luminal layer. We propose ACD spectroscopy to be a valuable tool linking the three-dimensional structure and the photophysical properties of pigment–protein complexes.	Jan 2019

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