Photophysics in single light-harvesting complexes II: from micelle to native nanodisks

J.M. Gruber, S. Scheidelaar, H. van Roon, J.P. Dekker, J.A. Kilian, R. van Grondelle

Research output: Chapter in Book / Report / Conference proceedingConference contributionAcademicpeer-review

Abstract

Most photosynthetic pigment-protein complexes of algae and higher plants are integral membrane proteins and are thus usually isolated in the presence of detergent to provide a hydrophobic interface and prevent aggregation. It was recently shown that the styrene maleic acid (SMA) copolymer can be used instead to solubilize and isolate protein complexes with their native lipid environment into nanodisk particles. We isolated LHCII complexes in SMA and compared their photophysics with trimeric LHCII complexes in β-DM detergent micelles to understand the effect of the native environment on the function of light-harvesting antennae. The triplet state kinetics and the calculated relative absorption cross section of single complexes indicate the successful isolation of trimeric complexes in SMA nanodisks, confirming the trimeric structure as the likely native configuration. The survival time of complexes before they photobleach is increased in SMA compared to detergent which might be explained by a stabilizing effect of the co-purified lipids in nanodisks. We furthermore find an unquenched fluorescence lifetime of 3.5 ns for LHCII in SMA nanodisks which coincides with detergent isolated complexes and notably differs from 2 ns typically found in native thylakoid structures. A large dynamic range of partially quenched complexes both in detergent micelles and lipid nanodisks is demonstrated by correlating the fluorescence lifetime with the intensity and likely reflects the conformational freedom of these complexes. This further supports the hypothesis that fluorescence intermittency is an intrinsic property of LHCII that may be involved in excess energy dissipation in native light-harvesting.
Original languageEnglish
Title of host publicationProc. of SPIE
Pages97140A
DOIs
Publication statusPublished - 2016

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Styrene
Micelles
Detergents
Fluorescence
Lipids
Algae
Pigments
Energy dissipation
Membrane Proteins
Proteins
Agglomeration
Copolymers
maleic acid
Antennas
Kinetics

Cite this

Gruber, J.M. ; Scheidelaar, S. ; van Roon, H. ; Dekker, J.P. ; Kilian, J.A. ; van Grondelle, R. / Photophysics in single light-harvesting complexes II: from micelle to native nanodisks. Proc. of SPIE. 2016. pp. 97140A
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abstract = "Most photosynthetic pigment-protein complexes of algae and higher plants are integral membrane proteins and are thus usually isolated in the presence of detergent to provide a hydrophobic interface and prevent aggregation. It was recently shown that the styrene maleic acid (SMA) copolymer can be used instead to solubilize and isolate protein complexes with their native lipid environment into nanodisk particles. We isolated LHCII complexes in SMA and compared their photophysics with trimeric LHCII complexes in β-DM detergent micelles to understand the effect of the native environment on the function of light-harvesting antennae. The triplet state kinetics and the calculated relative absorption cross section of single complexes indicate the successful isolation of trimeric complexes in SMA nanodisks, confirming the trimeric structure as the likely native configuration. The survival time of complexes before they photobleach is increased in SMA compared to detergent which might be explained by a stabilizing effect of the co-purified lipids in nanodisks. We furthermore find an unquenched fluorescence lifetime of 3.5 ns for LHCII in SMA nanodisks which coincides with detergent isolated complexes and notably differs from 2 ns typically found in native thylakoid structures. A large dynamic range of partially quenched complexes both in detergent micelles and lipid nanodisks is demonstrated by correlating the fluorescence lifetime with the intensity and likely reflects the conformational freedom of these complexes. This further supports the hypothesis that fluorescence intermittency is an intrinsic property of LHCII that may be involved in excess energy dissipation in native light-harvesting.",
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Photophysics in single light-harvesting complexes II: from micelle to native nanodisks. / Gruber, J.M.; Scheidelaar, S.; van Roon, H.; Dekker, J.P.; Kilian, J.A.; van Grondelle, R.

Proc. of SPIE. 2016. p. 97140A.

Research output: Chapter in Book / Report / Conference proceedingConference contributionAcademicpeer-review

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AU - Scheidelaar, S.

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AU - Kilian, J.A.

AU - van Grondelle, R.

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