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

Hancock, Ashley M.; Son, Minjung; Nairat, Muath; Wei, Tiejun; Jeuken, Lars J. C.; Duffy, Christopher D. P.; Schlau‐Cohen, Gabriela S.; Adams, Peter G.

doi:10.1039/d1cp01628h

Cited by 11 publications

(13 citation statements)

References 91 publications

(137 reference statements)

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“…Overall, these results show that the degree of quenching achieved with in-membrane electrophoresis can be controlled by simple modifications to the starting composition of the membrane. This could allow this platform to be used to investigate quenching over a wide dynamic range of concentrations for many other electrostatically charged fluorophores, such as different organic molecules or even membrane proteins, such as the light-harvesting pigment-protein complexes involved in photosynthesis in plants and bacteria. ,,, …”

Section: Resultsmentioning

confidence: 99%

“…This could allow this platform to be used to investigate quenching over a wide dynamic range of concentrations for many other electrostatically charged fluorophores, such as different organic molecules or even membrane proteins, such as the light-harvesting pigmentprotein complexes involved in photosynthesis in plants and bacteria. 38,39,45,46 The limitations of the electrophoresis technique and the analysis methodology should be briefly discussed in a wider context. First, the electrophoresis technique has the requirement that the fluorescent probe must respond to an electric field; therefore, alternative lipid-linked molecules that have either (single or multiple) negative or positive charges are likely to be effective, but neutral molecules will not undergo electrophoresis.…”

Section: Consistency Of Concentration Profiles Across Multiple Membra...mentioning

confidence: 99%

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Self-Quenching Behavior of a Fluorescent Probe Incorporated within Lipid Membranes Explored Using Electrophoresis and Fluorescence Lifetime Imaging Microscopy

et al. 2023

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Fluorescent probes are useful in biophysics research to assess the spatial distribution, mobility, and interactions of biomolecules. However, fluorophores can undergo “self-quenching” of their fluorescence intensity at high concentrations. A greater understanding of concentration-quenching effects is important for avoiding artifacts in fluorescence images and relevant to energy transfer processes in photosynthesis. Here, we show that an electrophoresis technique can be used to control the migration of charged fluorophores associated with supported lipid bilayers (SLBs) and that quenching effects can be quantified with fluorescence lifetime imaging microscopy (FLIM). Confined SLBs containing controlled quantities of lipid-linked Texas Red (TR) fluorophores were generated within 100 × 100 μm corral regions on glass substrates. Application of an electric field in-plane with the lipid bilayer induced the migration of negatively charged TR-lipid molecules toward the positive electrode and created a lateral concentration gradient across each corral. The self-quenching of TR was directly observed in FLIM images as a correlation of high concentrations of fluorophores to reductions in their fluorescence lifetime. By varying the initial concentration of TR fluorophores incorporated into the SLBs from 0.3% to 0.8% (mol/mol), the maximum concentration of fluorophores reached during electrophoresis could be modulated from 2% up to 7% (mol/mol), leading to the reduction of fluorescence lifetime down to 30% and quenching of the fluorescence intensity down to 10% of their original levels. As part of this work, we demonstrated a method for converting fluorescence intensity profiles into molecular concentration profiles by correcting for quenching effects. The calculated concentration profiles have a good fit to an exponential growth function, suggesting that TR-lipids can diffuse freely even at high concentrations. Overall, these findings prove that electrophoresis is effective at producing microscale concentration gradients of a molecule-of-interest and that FLIM is an excellent approach to interrogate dynamic changes to molecular interactions via their photophysical state.

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Section: Resultsmentioning

confidence: 99%

Section: Consistency Of Concentration Profiles Across Multiple Membra...mentioning

confidence: 99%

Self-Quenching Behavior of a Fluorescent Probe Incorporated within Lipid Membranes Explored Using Electrophoresis and Fluorescence Lifetime Imaging Microscopy

et al. 2023

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“…Our previous work has shown that plant LHCII [40, 41] can be “enhanced” with lipid-linked Texas Red (TR) chromophores when they are co-assembled into model membranes in the form of either lipid vesicles [36] or lipid nanodiscs [42]. This TR moiety is amphiphilic and is tethered to lipid headgroups which allow the possibility of close contact with membrane proteins ( Fig.…”

Section: Resultsmentioning

confidence: 99%

“…In these studies we demonstrated several advantages of using lipid-tagged chromophores: (i) the system is modular in the sense that dyes can be incorporated at a wide range of concentrations, (ii) LH proteins can be “enhanced” whilst in a biologically-relevant membrane environment, and (iii) membranes readily adsorb to surfaces making them amenable to surface-based nanotechnologies [36]. Evidence was presented for attractive interactions occurring between the TR chromophores and LHCII proteins which increased the rate of energy transfer [42]. For the current study, we hypothesized that it should be possible to enhance other LH complexes in a similar manner, and chose to study the LH2 antenna from Rhodobacter ( Rba. )…”

Section: Resultsmentioning

confidence: 99%

“…The drawback is the absence of direct control over the pigment-protein separation distance, leading to a continuum of distances from the lipid-dye to the protein. Consequently, the FRET efficiency will depend on the relative concentrations of each component and on the lateral diffusion of lipids [36, 65]. In summary, both types of coupling strategies can be effective and the choice may depend upon the nanomaterials desired by the researcher.…”

Section: Resultsmentioning

confidence: 99%

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Enhancing the spectral range of plant and bacterial Light-Harvesting pigment-protein complexes with various synthetic chromophores incorporated into lipid vesicles

Hancock

Swainsbury

Meredith

et al. 2022

Preprint

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The Light-Harvesting (LH) pigment-protein complexes found in photosynthetic organisms have the role of absorbing solar energy with high efficiency and transferring it to reaction centre complexes. LH complexes contain a suite of pigments that each absorb light at specific wavelengths, however, the natural combinations of pigments within any one protein complex do not cover the full range of solar radiation. Here, we provide an in-depth comparison of the relative effectiveness of five different organic "dye" molecules (Texas Red, ATTO, Cy7, DiI, DiR) for enhancing the absorption range of two different LH membrane protein complexes (the major LHCII from plants and LH2 from purple phototrophic bacteria). Proteoliposomes were self-assembled from defined mixtures of lipids, proteins and dye molecules and their optical properties were quantified by absorption and fluorescence spectroscopy. Both lipid-linked dyes and alternative lipophilic dyes were found to be effective excitation energy donors to LH protein complexes, without the need for direct chemical or generic modification of the proteins. The Forster theory parameters (e.g., spectral overlap) were compared between each donor-acceptor combination and found to be good predictors of an effective dye-protein combination. At the highest dye-to-protein ratios tested (over 20:1), the effective absorption strength integrated over the full spectral range was increased to ~180% of its natural level for both LH complexes. Lipophilic dyes could be inserted into pre-formed membranes although their effectiveness was found to depend upon favourable physicochemical interactions. Finally, we demonstrated that these dyes can also be effective at increasing the spectral range of surface-supported models of photosynthetic membranes, using fluorescence microscopy. The results of this work provide insight into the utility of self-assembled lipid membranes and the great flexibility of LH complexes for interacting with different dyes.

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Evidence for a transfer-to-trap mechanism of fluorophore concentration quenching in lipid bilayers

Meredith,

Kusunoki,

Evans

et al. 2024

Biophysical Journal

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

Abstract: We characterize the photophysical interactions between lipid-linked chromophores and plant light-harvesting proteins incorporated into nanodiscs using optical spectroscopy, simulations and theoretical modelling.

Cited by 11 publications

References 91 publications

Self-Quenching Behavior of a Fluorescent Probe Incorporated within Lipid Membranes Explored Using Electrophoresis and Fluorescence Lifetime Imaging Microscopy

Self-Quenching Behavior of a Fluorescent Probe Incorporated within Lipid Membranes Explored Using Electrophoresis and Fluorescence Lifetime Imaging Microscopy

Enhancing the spectral range of plant and bacterial Light-Harvesting pigment-protein complexes with various synthetic chromophores incorporated into lipid vesicles

Evidence for a transfer-to-trap mechanism of fluorophore concentration quenching in lipid bilayers

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