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Coupling of laser excitation and inelastic neutron scattering: attempt to probe the dynamics of light-induced C-phycocyanin dynamics

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Abstract

Excitation energy transfer (EET) in light-harvesting antennae is a highly efficient key event in photosynthesis, where light-induced dynamics of the antenna pigment-protein complexes may play a functional role. So far, however, the relationship between EET and protein dynamics remains unknown. C-phycocyanin (C-PC) is the main pigment/protein complex present in the cyanobacterial antenna, called “phycobilisome”. The aim of the present study was to investigate light-induced C-PC internal thermal motions (ps timescale) measured by inelastic neutron scattering. To synchronize the beginning of the laser flash (6 ns duration) with that of the neutron test pulse (∼87 μs duration), we developed a novel type of “time-resolved” experimental setup on MIBEMOL time-of-flight neutron spectrometer (LLB, France). Data acquisition has been modified to get quasi-simultaneously “light” and “dark” measurements (with and without laser, respectively) and eliminate many spurious effects that could occur on the sample during the experiment. The study was carried out on concentrated C-PC (∼135 g/L protein in D2O phosphate buffer), contained in an aluminium/sapphire sample holder (almost “transparent” for neutrons) and homogeneously illuminated inside an “integrating sphere”. We observed very similar incoherent dynamical structure factors of C-PC with or without light. The vibrational density of states showed two very slightly increased vibrational modes with light, at ∼30 and ∼50 meV (∼240 and ∼400 cm−1, respectively). These effects have to be verified by further experiments before probing any temporal evolution, by introducing a time delay between the laser flash and the neutron test pulse.

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Acknowledgments

We are very grateful to Dr. Alain Boussac (IBITEC-S, CEA-Saclay, France) for providing us with cyanobacterium phycobilisome extracts to purify C-PC. We also thank Dr. Marco Bonetti (IRAMIS/SPEC, CEA-Saclay, France) and Dr. Dominique Porterat (IRAMIS/SPAM, CEA-Saclay, France) for their expertise on the laser setup. J. P. gratefully acknowledges financial support by Deutsche Forschungsgemeinschaft (SFB 429, TP A1).

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Authors and Affiliations

  1. Laboratoire Léon-Brillouin UMR 12 CEA/CNRS, CEA-Saclay, 91191, Gif-sur-Yvette CEDEX, France

    Sophie Combet, Frédéric Coneggo, Jean-Pierre Ambroise, Marie-Claire Bellissent-Funel & Jean-Marc Zanotti

  2. Max-Volmer-Laboratories for Biophysical Chemistry, Technische Universität Berlin, Straße des 17, Juni 135, 10623, Berlin, Germany

    Jörg Pieper

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  1. Sophie Combet
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Correspondence to Sophie Combet.

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Advanced neutron scattering and complementary techniques to study biological systems. Contributions from the meetings, “Neutrons in Biology”, STFC Rutherford Appleton Laboratory, Didcot, UK, 11–13 July and “Proteins At Work 2007”, Perugia, Italy, 28–30 May 2007.

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Combet, S., Pieper, J., Coneggo, F. et al. Coupling of laser excitation and inelastic neutron scattering: attempt to probe the dynamics of light-induced C-phycocyanin dynamics. Eur Biophys J 37, 693–700 (2008). https://doi.org/10.1007/s00249-008-0320-1

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  • DOI: https://doi.org/10.1007/s00249-008-0320-1

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