Influence of detergent concentration on aggregation and spectroscopic properties of light-harvesting complex II |
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Authors: | Bernd Voigt Maria Krikunova Heiko Lokstein |
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Affiliation: | 1. Institut für Physik/Photonik, Universit?t Potsdam, Postfach 601553, 14415, Potsdam, Germany 2. Institut für Experimentalphysik, Universit?t Hamburg, Luruper Chaussee 149, 22761, Hamburg, Germany 3. Institut für Biochemie und Biologie/Pflanzenphysiologie, Universit?t Potsdam, Karl-Liebknecht-Str. 24-25, Haus 20, 14476, Potsdam-Golm, Germany
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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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Keywords: | Aggregation Laser spectroscopy LHC II Light-harvesting complex Nonlinear polarization spectroscopy in the frequency domain Non-photochemical quenching Pigment– pigment interactions |
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