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Photobleaching of chlorophyll in light-harvesting complex II increases in lipid environment

DOI: 10.3389/fpls.2020.00849 DOI Help

Authors: Monika Lingvay (Institute of Plant Biology, Biological Research Centre, Hungarian Academy of Sciences; University of Szeged) , Parveen Akhtar (Institute of Plant Biology, Biological Research Centre, Hungarian Academy of Sciences) , Krisztina Sebők-Nagy (Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences) , Tibor Páli (Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences) , Petar H. Lambrev (Institute of Plant Biology, Biological Research Centre, Hungarian Academy of Sciences)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Frontiers In Plant Science , VOL 11

State: Published (Approved)
Published: June 2020
Diamond Proposal Number(s): 17698

Open Access Open Access

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.

Journal Keywords: electron paramagnetic resonance; non-photochemical quenching; photoinhibition; photosystem II; reconstituted membranes; singlet oxygen

Diamond Keywords: Photosynthesis

Subject Areas: Biology and Bio-materials, Physics

Instruments: B23-Circular Dichroism

Added On: 23/07/2020 14:25


Discipline Tags:

Earth Sciences & Environment Plant science Life Sciences & Biotech Biophysics

Technical Tags:

Spectroscopy Circular Dichroism (CD)