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Influence of detergent concentration on aggregation and spectroscopic properties of light-harvesting complex II

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Abstract

Aggregation of photosynthetic light-harvesting complexes strongly influences their spectroscopic properties. Fluorescence yield and excited state lifetimes of the main light-harvesting complex (LHC II) of higher plants strongly depend on its aggregation state. Detergents are commonly used to solubilize membrane proteins and/or to circumvent their aggregation in aqueous environments. Nonlinear polarization spectroscopy in the frequency domain (NLPF) was performed with LHC II over a wide concentration range of the mild detergent n-dodecyl β-d-maltoside (β-DM). Additionally, conventional absorption-, fluorescence- and circular dichroism-spectra were measured.

The results indicate that: (i) conventional spectroscopic techniques are not well suited to investigate aggregation effects. NLPF provides a novel approach to overcome this problem: NLPF spectra display dramatic alterations upon even minor β-DM concentration changes. (ii) Commonly used detergent concentrations (around or slightly above the critical micellar concentration) apparently do not lead to complete trimerization of LHC II. A long-wavelength species in the NLPF spectra (peaking at about 685 nm), indicative of residual aggregation, persists up to DM-concentrations of 0.06%. (iii) High-resolution NLPF spectra indicate the existence of a species with a considerably shortened excited state lifetime. (iv) No indication of denaturation was found even at the highest β-DM concentrations used. (v) A specific change in interaction between certain chlorophyll(s) b and a xanthophyll molecule, probably neoxanthin, was detected upon aggregation as well as at higher β-DM concentrations. The results are discussed with respect to the still elusive mechanism of nonradiative dissipation of excess excitation energy in the antenna system.

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Abbreviations

β-DM:

n-Dodecyl β-d-maltoside

Chl:

Chlorophyll

CMC:

Critical micellar concentration

EET:

Excitation energy transfer

FWHM:

Full width at half-maximum

LHC II:

Light-harvesting complex II

NLPF:

Nonlinear polarization spectroscopy in the frequency domain

RT:

Room temperature

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Acknowledgements

Financial support by the Deutsche Forschungsgemeinschaft (SFB 429, TP A2) is gratefully acknowledged. The authors thank Prof. Dr. W. I. Gruszecki for stimulating discussions.

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Authors and Affiliations

  1. Institut für Physik/Photonik, Universität Potsdam, Postfach 601553, 14415, Potsdam, Germany

    Bernd Voigt

  2. Institut für Experimentalphysik, Universität Hamburg, Luruper Chaussee 149, 22761, Hamburg, Germany

    Maria Krikunova

  3. Institut für Biochemie und Biologie/Pflanzenphysiologie, Universität Potsdam, Karl-Liebknecht-Str. 24-25, Haus 20, 14476, Potsdam-Golm, Germany

    Heiko Lokstein

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  1. Bernd Voigt
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  2. Maria Krikunova
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  3. Heiko Lokstein
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Correspondence to Heiko Lokstein.

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Voigt, B., Krikunova, M. & Lokstein, H. Influence of detergent concentration on aggregation and spectroscopic properties of light-harvesting complex II. Photosynth Res 95, 317–325 (2008). https://doi.org/10.1007/s11120-007-9250-5

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  • DOI: https://doi.org/10.1007/s11120-007-9250-5

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