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Amphipols and Photosynthetic Light-Harvesting Pigment-Protein Complexes

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Abstract

The trimeric light-harvesting complexes II (LHCII) of plants and green algae are pigment-protein complexes involved in light harvesting and photoprotection. Different conformational states have been proposed to be responsible for their different functions. At present, detergent-solubilized LHCII is used as a model for the “light-harvesting conformation”, whereas the “quenched conformation” is mimicked by LHCII aggregates. However, none of these conditions seem to perfectly reproduce the properties of LHCII in vivo. In addition, several monomeric LHC complexes are not fully stable in detergent. There is thus a need to find conditions that allow analyzing LHCs in vitro in stable and, hopefully, more native-like conformations. Here, we report a study of LHCII, the major antenna complex of plants, in complex with amphipols. We have trapped trimeric LHCII and monomeric Lhcb1 with either polyanionic or non-ionic amphipols and studied the effect of these polymers on the properties of the complexes. We show that, as compared to detergent solutions, amphipols have a stabilizing effect on LHCII. We also show that the average fluorescence lifetime of LHCII trapped in an anionic amphipol is ~30 % shorter than in α-dodecylmaltoside, due to the presence of a conformation with 230-ps lifetime that is not present in detergent solutions.

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Abbreviations

A8-35:

A specific type of polyacrylate-based amphipol

AIBN:

Azobisisobutyronitrile

APol:

Amphipol

BR:

Bacteriorhodopsin

Car:

Carotenoid

CD:

Circular dichroism

Chl:

Chlorophyll

LHCII:

LHCII trimers

α-DDM:

N-dodecyl-α-d-maltoside

β-DDM:

N-dodecyl-β-d-maltoside

MD:

Molecular dynamics

MP:

Membrane protein

NA13:

A specific batch of non-ionic APol

NAPol:

Non-ionic APol

NPQ:

Non-photochemical quenching

SEC:

Size exclusion chromatography

TR:

Thiol-based transfer agent

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Acknowledgments

Particular thanks are due to Roberta Croce (VU University Amsterdam) for her support and help throughout this work, as well as for contributing to its writing. We also thank Laura M. Roy and Bart Sasbrink (VU University Amsterdam) for critical reading of the manuscript, and Pengqi Xu for his help with generating Fig. 7. This project was supported by the Chemical Science division of the Netherlands organization for scientific research (NWO-CW) via an ECHO grant to R. Croce, by the CNRS, by University Paris-7, and by the “Initiative d’Excellence” program from the French State (Grant “DYNAMO”, ANR-11-LABX-0011-01).

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Authors and Affiliations

  1. Department of Physics and Astronomy, Faculty of Sciences, VU University Amsterdam, De Boelelaan 1081, 1081 HV, Amsterdam, The Netherlands

    Milena Opačić

  2. Equipe Chimie Bioorganique et Systèmes Amphiphiles, Université d’Avignon, 33 rue Louis Pasteur, 84000, Avignon, France

    Grégory Durand, Michael Bosco & Ange Polidori

  3. Institut des Biomolécules Max Mousseron (UMR 5247), 15 Avenue Charles Flahault, 34093, Montpellier Cedex 05, France

    Grégory Durand, Michael Bosco & Ange Polidori

  4. Unité Mixte de Recherche 7099, Centre National de la Recherche Scientifique and Université Paris 7, Institut de Biologie Physico-Chimique (FRC 550), 13 rue Pierre-et-Marie Curie, 75005, Paris, France

    Jean-Luc Popot

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  1. Milena Opačić
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  2. Grégory Durand
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Correspondence to Milena Opačić.

Electronic Supplementary Material

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Supplementary information: Scheme of NA13 synthesis; size exclusion chromato-gra-phy profiles of A8-35-trapped Lcb1 and of Lhcb1 in α-DDM; fluorescence emission spectra of A8-35- and NA13-trapped LHCII and of LHCII kept in α-DDM; sucrose gradients analysis of LHCII trimers either NA13-trapped or kept in α-DDM, at pH 5.5, 6.5 and 7.6.

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Opačić, M., Durand, G., Bosco, M. et al. Amphipols and Photosynthetic Light-Harvesting Pigment-Protein Complexes. J Membrane Biol 247, 1031–1041 (2014). https://doi.org/10.1007/s00232-014-9712-6

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