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Michel Négrerie

Chercheur INSERM
Pièce : 02-2015

Tél :01 69 33 50 52
fax : 01 69 33 50 84

michel.negrerie at polytechnique.edu
                        

Research topics :


Dynamique structurale des senseurs du NO et de la  NO-synthase

The proteins investigated in this research are involved in the Nitric Oxide signal transduction pathway  :

Bacterial NO-sensors and Cytochrome c'

Mitochondrial Cytochrome c

This research aims at investigating :

  • the mechanisms at molecular level by which the binding of the effector NO activates the catalytic activity of guanylate cyclase.
  • how guanylate cyclase is deactivated.
  • the general rules defining how heme proteins control the dynamics of nitric oxide as a ligand.
  •  how NO-synthase controls the dynamic exchange between the substrate O2 and the product NO (i.e. to and from the heme) during continuous turn-over conditions, and how cofactors and substrates analogs affect these dynamics.

Whereas O2 is the substrate of NO-synthase, NO produced by NO-synthase is an effector of guanylate cyclase. Although not fully demonstrated, the binding of NO to cytochromes c and c' are thought to be physiologically relevant. Bacterial NO-sensors, which are homologous to the heme domain of guanylate have been recently discovered.

We studied also mutants of myoglobin as models to investigate the role of heme coordination in the binding and release of NO and in the signal transduction.

Our investigations on cytochrome c' from Alcaligenes xylosoxidans have shown that its dynamic properties correspond to those of a NO transporter protein.

Among several methodologies and techniques, our tools comprise time-resolved visible absorption spectroscopy (40 fs resolution, up to 5 ns) and time-resolved Raman spectroscopy (0.6 ps resolution, up to 1.5 ns).

 

Collaborations :


Dr Serguei Kruglik : Time-resolved Raman spectroscopy.

BioMoCeTi, CNRS UMR 7033, University Pierre et Marie Curie, Génopole Campus, 91030 Evry, France.

Dr Pierre Nioche : NO-sensors homologous to guanylate cyclase.

INSERM, UMR-S 747, Université Paris 5, France.

Pr Ah-Lim Tsai : Endothelial NO-Synthase.

Hematology and Internal Medicine, University of Texas Medical School, Houston, Texas, USA.

Pr Colin Andrew : Cytochrome c'.

Biochemistry and Molecular Biology, Institut of Sciences and Technology, Beaverton, Oregon, USA.

Pr C.S. Raman : NO-sensors homologous to guanylate cyclase.

Biochemistry and Molecular Biology, University of Texas Medical School, Houston, Texas, USA.

Pr Stefan Franzen : Structural dynamics of NO in myoglobin.

Department of Chemistry, North Carolina State University, Raleigh, NC 27695, USA.

 

Selected  publications :


Sauviat, M.-P., Colas, A., Chauveau, M.-J., Drapier, J.-C. &Negrerie, M."Hypericin activates L-type Ca2+ channels in cardiac myocytes" (2007) J. Nat. Prod. 70, 510-514.

Kruglik, S. G., Lambry, J.-C., Cianetti,  S., Martin, J.-L. , Eady, R. R., Andrew, C. R. & Négrerie, M."Molecular basis for nitric oxide dynamics and affinity with Alcaligenes xylosoxidans cytochrome c' " (2007) J. Biol. Chem. 282, 5053-5062.

M. Négrerie, S. G. Kruglik, J.-C. Lambry, M. H. Vos, J.-L. Martin & S. Franzen"Role of Heme Iron Coordination and Protein Structure in the Dynamics and Geminate Rebinding of Nitric Oxide to H93G Myoglobin : Implications for NO-Sensors" (2006) J. Biol. Chem. 281, 10389-10398.

M. Négrerie, S. Cianetti, M. H. Vos, J.-L. Martin & S. G. Kruglik"Photoinduced coordination dynamics of cytochrome c: ferrous versus ferric species studied by time-resolved resonance Raman and transient absorption spectroscopies" (2006) J. Phys. Chem. B.  110, 12766-12781.

M. Négrerie, J.-L. Martin & H.-O. Nghiêm "Functionality of nitrated acetylcholine receptor : the two-step formation of nitrotyrosines reveals their differential role in agonist and neurotoxin binding" (2005) FEBS Letters. 579, 2643-2647.

Ciannetti, M. Négrerie, M. H. Vos, J.-L. Martin, & S.G. Kruglik, "Photodissociation of heme distal methionine in ferrous cytochrome c revealed by sub-picosecond time-resolved resonance Raman spectroscopy" (2004) J. Am. Chem. Soc.126, 13932-13933.

Cianetti S., Kruglik, S.G., Vos, M.H., Turpin, P.-Y., Martin J.-L. & Négrerie, M."Interaction of cytochrome c with NO studied by time-resolved raman and absorption spectroscopy" (2004) Biochim. Biophys. Acta-Bionergetics1658, 219 Suppl. Proceedings of the 13th EBEC (2004 European BioEnergetics Conference) Pisa, Italy.

Gautier, C., Negrerie, M., Wang, Z.-Q., Lambry, J.-C., Stuehr, D. J., Collin, F., Martin, J.-L. & Slama-Schwok, A. "Dynamic Regulation of the Inducible Nitric-oxide Synthase by NO: comparaison with the endothelial isoform" (2004) J. Biol. Chem.279, 4358-4365.

Liebl, U., Bouzhir-Sima, L., Kiger, L., Marden, M. C., Lambry, J.C., Negrerie, M. & Vos, M.H. "Ligand binding dynamics to the heme domain of the oxygen sensor Dos from Escherichia coli" (2003) Biochemistry42, 6527-6535.

Chowdhury, P., Gondry, M., Genet, R., Martin, J.L., Ménez, A., Negrerie, M. & Petrich  J. W. Picosecond dynamics of a peptide from the acetylcholine receptor interacting with a neurotoxin probed by tailored tryptophan fluorescence" (2003) Photochem. Photobiol.77, 151-157.

Liebl, U., Bouzhir-Sima, L., Negrerie, M., Martin, J.-L. & Vos, M.H. "Ultrafast ligand rebinding in the heme domain of the oxygen sensors FixL ans Dos : general regulatory implications for heme-based sensors" (2002). Proc. Natl. Acad. Sci. USA. 99, 12771-12776.

Chirvony, V., Negrerie, M., Martin, J.-L. & Turpin, P.-Y. "Subpicosecond dynamics and mechanisms of quenching of photoexcited Cu(II)-porphyrin in solutions of Lewis bases" (2002). J. Phys. Chem.106, 5760-5767.

Slama-Schwok, A., Negrerie, M., Berka, V., Lambry, J.C., Tsai, A.L., Vos, M.H. & Martin, J.L. "Nitric oxide traffic in endothelial NO-synthase: evidence for a new NO binding site dependent on tetrahydrobiopterin?" (2002). J. Biol. Chem., 277, 7581-7586.

Negrerie, M., Bouzhir, L., Martin, J.-L. & Liebl, U. "Control of nitric oxide dynamics by guanylate cyclase in its activated state" (2001). J. Biol. Chem., 276, 46815-46821.

Liebl, U, Lipowski, G., Negrerie, M., Lambry, J.-C., Martin, J.-L. & Vos, M.H. "Coherent reaction dynamics in a bacterial cytochrome c oxidase" (1999) Nature 401, 181-184.

Négrerie, M., Berka, V., Vos, M.H., Liebl, U., Lambry, J.-C., Tsai, A-L., & Martin, J.-L. "Geminate recombination of nitric oxide to endothelial nitric oxide-synthase and mechanistic implications" (1999) J. Biol. Chem.274, 24694-24702