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FRENCH INFRASTRUCTURE FOR INTEGRATED STRUCTURAL BIOLOGY

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SOUTH PARIS

ELECTRONIC SPECTROSCOPY

The Service de Bioénergétique, Biologie Structurale et Mécanismes(SB2SM) / CNRS-UMR9198, located at JOLIOT/I2BC/CEA-Saclay, has a platform dedicated to the electronic spectroscopy of biological samples.  Historically, we specialised in the study of photosynthesis/photophysics and over the years have accumulated a diverse range of commercially available spectrometers as well as home-built optical set-ups including a “Machine de Joliot”. In general, we are able to monitor spectroscopic changes at the protein level in intact cells on many time scales for research projects that wished to monitor absorption/fluorescence changes.

A special set-up1-3 was developed to monitor small flash-induced transient absorption changes with a time resolution of 300 ps in single flash experiments, or with a very modest number of averaged experiments.

Commercial spectrometers are available to measure chlorophyll fluorescence, P700 oxidation and NADPH fluorescence. Chlorophyll fluorescence and P700 can be measuredin vivousing leaves or algae/cyanobacteria cultures. NADPH fluorescence can be measured in algae/cyanobacteria cultures or in isolated intact chloroplasts.

Thermoluminescence can be measured with a home-built set-up that is able to monitor charge recombination reactions in photosystem II. This method can be used for leaves, algae/cyanobacteria cultures as well as isolated photosytem II.

The platform consists of:

Bibliography

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  2. Thiagarajan V., Byrdin M., Eker A.P.M, Müller  P. and Brettel K. (2011) Kinetics of cyclobutane thymine dimer splitting by DNA photolyase directly monitored in the UV. Proc. Natl. Acad. Sci. USA 108: 9402-9407.
  3. Müller P. and Brettel K. (2012) [Ru(bpy)3]2+ as a reference in transient absorption spectroscopy: differential absorption coefficients for formation of the long-lived 3MLCT excited state. Photochem. Photobiol. Sci. 11: 632-636.
  4. Müller P., Yamamoto J., Martin R., Iwai S. and Brettel K. (2015) Discovery and functional analysis of a 4th electron-transferring tryptophan conserved exclusively in animal cryptochromes and (6-4) photolyases, Chem. Commun., 51, 85: 15502-15505.
  5. Müller P., Brettel K., Grama L., Nyitrai M. and  Lukacs A. (2016) Photochemistry of Wild-Type and N378D Mutant E. coli DNA Photolyase with Oxidized FAD Cofactor Studied by Transient Absorption Spectroscopy, Chemphyschem, 17, 9: 1329-1340.
  6. Yamamoto J., Plaza P. and Brettel K. (2017) Repair of (6-4) Lesions in DNA by (6-4) Photolyase: 20 Years of Quest for the Photoreaction Mechanism, Photochemistry and Photobiology, 93, 1: 51-66.
  7. Roach T., Sedoud A. and Krieger-Liszkay  A. (2013) Acetate in mixotrophic growth medium affects photosystem II in Chlamydomonas reinhardtii and protects against photoinhibition. Biochim. Biophys. Acta 1827: 1183-1890.
  8. Galzerano D., Feilke K., Schaub P., Beyer P. and Krieger-Liszkay A. (2013) Effect of constitutive expression of bacterial desaturase CRTI on photosynthetic electron transport in Arabidopsis thaliana. Biochim. Biophys. Acta 1837: 345-353.
  9. Gwizdala M., Wilson A. and Kirilovsky  D. (2011) In vitro reconstitution of the cyanobacterial photoprotective mechanism mediated by the Orange Carotenoid Protein. Plant Cell 23: 2631-2643.
  10. Guerrero F., Sedoud A., Kirilovsky D., Rutherford W.A., Ortega J.M. and Roncel M. (2011) A high redox potential form of cytochrome c550 in Photosystem II from Thermosynechococcus elongates.  J. Biol. Chem. 286: 5985-5994.
  11. Sétif P., Hirasawa M., Cassan N., Lagoutte B., Tripathy J.N. and  Knaff DB (2009) New insights into the catalytic cycle of plant nitrite reductase. Electron transfer kinetics and charge storage. Biochemistry 48: 2828-2838.
  12. Korn A., Ajlani G., Lagoutte B., Gall A., and Sétif P (2009) Ferredoxin:NADP+ oxidoreductase association with phycocyanin modulates its properties. J. Biol. Chem. 284: 31789-31797.
  13. Trinkunas G., Zerlauskiene O., Urboniene V., Chmeliov  J., Gall A., Robert B. and Valkunas L. (2012) Exciton band structure in bacterial peripheral light-harvesting complexes. J. Phys. Chem. B 116: 5192-5198.
  14. Biniek C., Heyno E., Kruk J., Sparla F., Trost P. and Krieger-Liszkay A.(2017) Role of the NAD(P)H quinone oxidoreductase NQR and the cytochrome b AIR12 in controlling superoxide generation at the plasma membrane, Planta, 245, 4:807-817. 
  15. Krieger-Liszkay A. and Feilke K. (2015),  The Dual Role of the Plastid Terminal Oxidase PTOX: Between a Protective and a Pro-oxidant Function , Frontiers in Plant Science, 6:1147.
  16. López-Igual R., Wilson A., Leverenz R. L., Melnicki M. R., Bourcier de Carbon C., Sutter M., Turmo A., Perreau F., Kerfeld C. A., and Kirilovsky D. (2016) Different Functions of the Paralogs to the N-Terminal Domain of the Orange Carotenoid Protein in the Cyanobacterium Anabaena sp. PCC 7120 , Plant Physiology, 171, 3: 1852-1866.
  17. Thurotte A., Bourcier de Carbon C., Wilson A., Talbot L., Cot S., López-Igual and Kirilovsky (2017) The cyanobacterial Fluorescence Recovery Protein has two distinct activities: Orange Carotenoid Protein amino acid involved in FRP interaction Biochimica et Biophysica Acta (BBA) - Bioenergetics, 1858, 4: 308-317.
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  19. Kauny  J. and  Sétif P. (2014) NADPH fluorescence in the cyanobacterium Synechocystis sp. PCC 6803: A versatile probe for in vivo measurements of rates, yields and pools. Biochim. Biophys. Acta, 1837,6: 792-801.
  20. Gall A., Ilioaia C., Kruger T.P.J., Novoderezhkin V.I., Robert B. and Grondelle, R. van (2015) Conformational Switching in a Light-Harvesting Protein as Followed by Single-Molecule Spectroscopy. Biophys. J. 108: 2713-2720.
  21. Chmeliov J., Songaila, E. Rancova O., Gall A., Robert B., Abramavicius D. and Valkunas L. (2013) Excitons in the LH3 Complexes from Purple Bacteria. J. Phys. Chem. B. 117: 11058-11068
  22. M.M. Mendes-Pinto, E. Sansiaume, H. Hashimoto, A.A. Pascal, A. Gall and Robert B. (2013) Electronic Absorption and Ground State Structure of Carotenoid Molecules, J. Phys. Chem. B. 117: 11015-11021.