Excitation quenching in chlorophyll-carotenoid antenna systems: 'coherent' or 'incoherent'

Plants possess an essential ability to rapidly down-regulate light-harvesting in response to high light. This photoprotective process involves the formation of energy-quenching interactions between the chlorophyll and carotenoid pigments within the antenna of Photosystem II (PSII). The nature of these interactions is currently debated, with, among others, ‘incoherent’ or ‘coherent’ quenching models (or a combination of the two) suggested by a range of time-resolved spectroscopic measurements. In ‘incoherent quenching’, energy is transferred from a chlorophyll to a carotenoid and is dissipated due to the intrinsically short excitation lifetime of the latter. ‘Coherent quenching’ would arise from the quantum mechanical mixing of chlorophyll and carotenoid excited state properties, leading to a reduction in chlorophyll excitation lifetime. The key parameters are the energy gap, Δ=Car−Chl, Δ ε = ε C a r − ε C h l , and the resonance coupling, J, between the two excited states. Coherent quenching will be the dominant process when −<Δ<, − J < Δ ε < J , i.e., when the two molecules are resonant, while the quenching will be largely incoherent when Chl>(Car+). ε C h l > ( ε C a r + J ) . One would expect quenching to be energetically unfavorable for Chl<(Car−). ε C h l < ( ε C a r − J ) . The actual dynamics of quenching lie somewhere between these limiting regimes and have non-trivial dependencies of both J and Δ. Δ ε . Using the Hierarchical Equation of Motion (HEOM) formalism we present a detailed theoretical examination of these excitation dynamics and their dependence on slow variations in J and Δ. Δ ε . We first consider an isolated chlorophyll–carotenoid dimer before embedding it within a PSII antenna sub-unit (LHCII). We show that neither energy transfer, nor the mixing of excited state lifetimes represent unique or necessary pathways for quenching and in fact discussing them as distinct quenching mechanisms is misleading. However, we do show that quenching cannot be switched ‘on’ and ‘off’ by fine tuning of Δ Δ ε around the resonance point, Δ=0. Δ ε = 0. Due to the large reorganization energy of the carotenoid excited state, we find that the presence (or absence) of coherent interactions have almost no impact of the dynamics of quenching. Counter-intuitively significant quenching is present even when the carotenoid excited state lies above that of the chlorophyll. We also show that, above a rather small threshold value of >10cm−1 J > 10 c m − 1 quenching becomes less and less sensitive to J (since in the window −<Δ< − J < Δ ε < J the overall lifetime is independent of it). The requirement for quenching appear to be only that >0. J > 0. Although the coherent/incoherent character of the quenching can vary, the overall kinetics are likely robust with respect to fluctuations in J and Δ. Δ ε . This may be the basis for previous observations of NPQ with both coherent and incoherent features

Unravelling the fluorescence kinetics of light-harvesting proteins with simulated measurements

The plant light-harvesting pigment-protein complex LHCII is the major antenna sub-unit of PSII and is generally (though not universally) accepted to play a role in photoprotective energy dissipation under high light conditions, a process known Non-Photochemical Quenching (NPQ). The underlying mechanisms of energy trapping and dissipation within LHCII are still debated. Various proposed models differ considerably in their molecular and kinetic detail, but are often based on different interpretations of very similar transient absorption measurements of isolated complexes. Here we present a simulated measurement of the fluorescence decay kinetics of quenched LHCII aggregates to determine whether this relatively simple measurement can discriminate between different potential NPQ mechanisms. We simulate not just the underlying physics (excitation, energy migration, quenching and singlet-singlet annihilation) but also the signal detection and typical experimental data analysis. Comparing this to a selection of published fluorescence decay kinetics we find that: (1) Different proposed quenching mechanisms produce noticeably different fluorescence kinetics even at low (annihilation free) excitation density, though the degree of difference is dependent on pulse width. (2) Measured decay kinetics are consistent with most LHCII trimers becoming relatively slow excitation quenchers. A small sub-population of very fast quenchers produces kinetics which do not resemble any observed measurement. (3) It is necessary to consider at least two distinct quenching mechanisms in order to accurately reproduce experimental kinetics, supporting the idea that NPQ is not a simple binary switch switch

Trivial Excitation Energy Transfer to Carotenoids Is an Unlikely Mechanism for Non-photochemical Quenching in LHCII

Higher plants defend themselves from bursts of intense light via the mechanism of Non-Photochemical Quenching (NPQ). It involves the Photosystem II (PSII) antenna protein (LHCII) adopting a conformation that favors excitation quenching. In recent years several structural models have suggested that quenching proceeds via energy transfer to the optically forbidden and short-lived S(1) states of a carotenoid. It was proposed that this pathway was controlled by subtle changes in the relative orientation of a small number of pigments. However, quantum chemical calculations of S(1) properties are not trivial and therefore its energy, oscillator strength and lifetime are treated as rather loose parameters. Moreover, the models were based either on a single LHCII crystal structure or Molecular Dynamics (MD) trajectories about a single minimum. Here we try and address these limitations by parameterizing the vibronic structure and relaxation dynamics of lutein in terms of observable quantities, namely its linear absorption (LA), transient absorption (TA) and two-photon excitation (TPE) spectra. We also analyze a number of minima taken from an exhaustive meta-dynamical search of the LHCII free energy surface. We show that trivial, Coulomb-mediated energy transfer to S(1) is an unlikely quenching mechanism, with pigment movements insufficiently pronounced to switch the system between quenched and unquenched states. Modulation of S(1) energy level as a quenching switch is similarly unlikely. Moreover, the quenching predicted by previous models is possibly an artifact of quantum chemical over-estimation of S(1) oscillator strength and the real mechanism likely involves short-range interaction and/or non-trivial inter-molecular states

Photosynthesis Under a Red Sun: Predicting the absorption characteristics of an extraterrestrial light-harvesting antenna

Here we discuss the feasibility of photosynthesis on Earth-like rocky planets in close orbit around ultra-cool red dwarf stars. Stars of this type have very limited emission in the \textit{photosynthetically active} region of the spectrum (400−700 nm), suggesting that they may not be able to support oxygenic photosynthesis. However, photoautotrophs on Earth frequently exploit very dim environments with the aid of highly structured and extremely efficient antenna systems. Moreover, the anoxygenic photosynthetic bacteria, which do not need to oxidize water to source electrons, can exploit far red and near infrared light. Here we apply a simple model of a photosynthetic antenna to a range of model stellar spectra, ranging from ultra-cool (2300 K) to Sun-like (5800 K). We assume that a photosynthetic organism will evolve an antenna that maximizes the rate of energy input while also minimizing fluctuations. The latter is the 'noise cancelling' principle recently reported by Arp et al. 2020. Applied to the Solar spectrum this predicts optimal antenna configurations in agreement with the chlorophyll Soret absorption bands. Applied to cooler stars, the optimal antenna peaks become redder with decreasing stellar temperature, crossing to the typical wavelength ranges associated with anoxygenic photoautotrophs at ∼3300 K. Lastly, we compare the relative input power delivered by antennae of equivalent size around different stars and find that the predicted variation is within the same order of magnitude. We conclude that low-mass stars do not automatically present light-limiting conditions for photosynthesis but they may select for anoxygenic organisms

Structural Basis for Allosteric Regulation in the Major Antenna Trimer of Photosystem II

The allosteric regulation of protein function proves important in many life-sustaining processes. In plant photosynthesis, LHCII, the major antenna complex of Photosystem II, employs a delicate switch between light harvesting and photoprotective modes. The switch is triggered by an enlarged pH gradient (ΔpH) across the thylakoid membranes. Using molecular simulations and quantum calculations, we show that ΔpH can tune the light-harvesting potential of the antenna via allosteric regulation of the excitonic coupling in chlorophyll-carotenoid pairs. To this end, we propose how the LHCII excited state lifetime is coupled to the environmental conditions. In line with experimental findings, our theoretical model provides crucial evidence toward the elucidation of the photoprotective switch of higher plants at an all-atom resolution

Ultrafast energy transfer between lipid-linked chromophores and plant light-harvesting complex II

Light-Harvesting Complex II (LHCII) is a membrane protein found in plant chloroplasts that has the crucial role of absorbing solar energy and subsequently performing excitation energy transfer to the reaction centre subunits of Photosystem II. LHCII provides strong absorption of blue and red light, however, it has minimal absorption in the green spectral region where solar irradiance is maximal. In a recent proof-of-principle study, we enhanced the absorption in this spectral range by developing a biohybrid system where LHCII proteins together with lipid-linked Texas Red (TR) chromophores were assembled into lipid membrane vesicles. The utility of these systems was limited by significant LHCII quenching due to protein–protein interactions and heterogeneous lipid structures. Here, we organise TR and LHCII into a lipid nanodisc, which provides a homogeneous, well-controlled platform to study the interactions between TR molecules and single LHCII complexes. Fluorescence spectroscopy determined that TR-to-LHCII energy transfer has an efficiency of at least 60%, resulting in a 262% enhancement of LHCII fluorescence in the 525–625 nm range, two-fold greater than in the previous system. Ultrafast transient absorption spectroscopy revealed two time constants of 3.7 and 128 ps for TR-to-LHCII energy transfer. Structural modelling and theoretical calculations indicate that these timescales correspond to TR–lipids that are loosely- or tightly-associated with the protein, respectively, with estimated TR-to-LHCII separations of ∼3.5 nm and ∼1 nm. Overall, we demonstrate that a nanodisc-based biohybrid system provides an idealised platform to explore the photophysical interactions between extrinsic chromophores and membrane proteins with potential applications in understanding more complex natural or artificial photosynthetic systems