CGM - Département Biologie Cellulaire et Intégrative
Ingénierie des Protéines Membranaires (LIPM)
Responsable : Denis POMPON
MàJ : 15/07/10
L'équipe
Louise Aigrain, Doctorante
Florence Calvayrac, Ingénieur
Fataneh Fatemi, Doctorante
Nathalie Franche, Etudiante Master 2
Aude Laisné, Ingénieur
Jacqueline Loeper, Maître de Conférences, UPMC
Denis Pompon, Directeur de Recherche, CNRS
Gilles Truan, Chargé de Recherche, CNRS
Philippe Urban, Chargé de Recherche, CNRS
Notre adresse
CNRS-CGM
Avenue de la Terrasse - Bât. 23B
91198 GIF-SUR-YVETTE Cedex
FRANCE
Téléphone : 33 (0)1 69 82 36 80
Fax : 33 (0)1 69 82 36 82
Thématiques
Le laboratoire entretient deux axes de recherche complémentaires, l'ingénierie métabolique et l'ingénierie moléculaire.
Ingénierie métabolique : forte de son passé, reconnue internationalement, pour l'expression hétérologue de protéines membranaires chez S. cerevisiae et au travers d'une longue collaboration avec des partenaires industriels (Hoechst Marion Roussel puis Aventis), notre équipe a reconstitué des voies de synthèse d'hormones stéroïdes chez la levure de boulangerie. Ce travail a nécessité la délétion et la modification de plusieurs gènes de la levure, pour détourner la voie de synthèse des stérols mais aussi l'introduction de plusieurs gènes provenant d'organismes différents pour réaliser les étapes de synthèse manquantes chez la levure.
Ingénierie protéique : notre équipe s'intéresse particulièrement aux relations structure-fonction chez les cytochromes P450s, enzymes qui participent à un complexe multienzymatique membranaire. Nous étudions notamment les différents modes de reconnaissance des substrats, des partenaires polypeptidiques, le rôle de la membrane dans la dynamique moléculaire et l'assemblage de ces complexes. Ces études sont réalisées à la fois avec des outils moléculaires (mutagénèse) et biochimiques (spectroscopie, cinétique rapide, etc.). Nous avons par ailleurs développé des stratégies d'évolution combinatoire pour mieux comprendre les mécanismes moléculaires qui permettent de générer une nouvelle fonction dans une protéine.
Publications (depuis 1997)
Nunez, M., Guittet, E., Pompon, D.,van Heijenoort, C., Truan, G. (2010) NMR structure note: oxidized microsomal human cytochrome b5. J Biomol NMR. 47 (7) 289-95.
Laisné, A., Pompon, D., Leroy, J.-L. (2010) [C7GC4]4 Association into supra molecular i-motif structures. Nucleic Acids Res, Epub ahead of print.
Fon Tacer, KF, Pompon, D, Rozman, D. (2010) Adaptation of cholesterol synthesis to fasting and TNF-alpha: Profiling cholesterol intermediates in the liver, brain, and testis. J Steroid Biochem Mol Biol, Epub ahead of print.
Panchal, M., Loeper, J., Cossec, J., Perruchini, C., Lazar, A., Pompon, D. and Duyckaerts, C. (2010) Enrichment of cholesterol in microdissected Alzheimer disease senile plaques as assessed by mass spectrometry. J Lipid Res, 51 (3) 598-605..
Kastelic, D., Frković-Grazio, S., Baty, D.,Truan, G., Komel, R. and Pompon, D. (2009) A single-step procedure for the selection of efficiently produced llama VH binders directed against cancer markers. J Immun Methods, 350 (1-2) 54-62.
Duval, A., Laisné, A., Pompon, D., Held, S., Bellemain, A., Moreau, J. and Canva, M. (2009) Polarimetric surface plasmon resonance imaging biosensor. Opt Lett, 34 (23) 3634-6.
Aigrain, L., Pompon, D., Moréra, S., Truan, G. (2009) Structure of the open conformation of a functional chimeric NADPH cytochrome P450 reductase. EMBO Rep, 10 (7) 742-7.
Nouet, C., Truan, G., Mathieu, L. and Dujardin, G. (2009) Functional Analysis of Yeast bcs1 Mutants Highlights the Role of Bcs1p-Specific Amino Acids in the AAA Domain. J Mol Biol, 388 (2) 252-61.
Aigrain, L., Pompon, D., Truan, G. and Moréra, S. (2009) Cloning, purification, crystallization and preliminary X-ray analysis of a chimeric NADPH-cytochrome P450 reductase. Acta Crystallogr Sect F Struct Biol Cryst Commun, 65 (Pt3) 210-2.
Jézéquel, L., Loeper, J. and Pompon, D. (2008) Sequence-independent construction of ordered combinatorial libraries with predefined crossover points. Biotechniques, 45 (5) 523-32.
Parker, J., Merkamm, M., Manning, N.-J., Pompon, D., Kelly, S.-L. and Kelly, D.-E. (2008) Differential azole antifungal efficacy contrasted using a yeast strain humanised for sterol 14{alpha}-demethylase at the homologous locus. Antimicrob Agents Chemother, 52 (10) 3597-603.
Acimovic, J., Fink, M., Pompon, D., Bjorkhem, I., Hirayama, J., Sassone-Corsi, P., Golicnik, M. and Rozman, D. (2008) CREM modulates the circadian expression of CYP51, HMGCR, and cholesterogenesis in the liver. Biochem Biophys Res Commun, 376 (1) 206-10.
Urban, P., Truan, G. and Pompon, D. (2008) High-throughput enzymology and combinatorial mutagenesis for mining cytochrome P450 functions. Expert Opin Drug Metab Toxicol., 4 (6) 733-747.
Pompon, D., Truan, G. and Urban, P. (2008) Ingénierie des cytochrome P450. Biofutur, 288 34-38.
Rezen, T., Juvan, P., Fon Tacer, K., Kuzman, D., Roth, A., Pompon, D., Aggerbeck, LP., Meyer, UA., Rozman, D. (2008) The Sterolgene v0 cDNA microarray: a systemic approach to studies of cholesterol homeostasis and drug metabolism. BMC Genomics 11;9:76.
Fon Tacer, K., Kuzman, D., Seliskar, M., Pompon, D. and Rozman, D. (2007) TNF-{alpha} interferes with lipid homeostasis and activates acute and pro-atherogenic processes Physiol Genomics, 31 (2) 216-227.
Pompon, D. and Laisné, A. (2007) PDNA as building blocks for membrane-guided self-assemblies. Biochem Soc Trans, 35 (Pt 3) 495-7.
Taly, V., Urban, P., Truan, G. and Pompon, D. (2007) A combinatorial approach to substrate discrimination in the P450 CYP1A subfamily. Biochim Biophys Acta (General Subjects), 1770 (3) 446-57.
Keita, B., Mbomekalle, I. M., de Oliveira, P., Ranjbari, A., Justum, Y., Nadjo, L. and Pompon, D. (2006) Reactions of 5-substituted polyoxometalates with cystein. Journal of Cluster Science, Apr 2006, 1 - 13. Pre publication on line
Muller, C., Pompon, D., Urban, P. and Morfin, R. (2006) Inter-conversion of 7alpha- and 7beta-hydroxy-dehydroepiandrosterone by the human 11beta-hydroxysteroid dehydrogenase type 1. J Steroid Biochem Mol Biol, 99 215-222.
Boireau, W., Duncan, A. and Pompon, D. (2005) Bioengineering and characterization of DNA-protein assemblies floating on supported membranes. in Methods in Molecular Biology 300, T. Vo-Dinh (Ed.), Humana Press, Totowa, NJ.
Boireau, W., Zeeh, J. C., Puig, P. E. and Pompon, D. (2005) Unique supramolecule asembly of a redox protein with nucleic acids onto hybrid bilayer: toward a dynamic DNA chip. Biosensors and Bioelectronics, 15 (20) 1631-7.
Kim, S., Chalbot, S., Pompon, D., Jo, D. and Morfin, R. (2004) The human cytochrome P4507B1: catalytic activity studies. J Steroid Biochem Mol Biol, 95 (5) 383-9.
Rouillard, J. M., Lee, W., Truan, G., Gao, X., Zhou, X., and Gulari, E. (2004). Gene2Oligo: oligonucleotide design for in vitro gene synthesis. Nucleic Acids Res 32, W176-180
Nthangeni, M., Urban, P., Pompon, D., Smit, M., and Nicaud, J. (2004). The use of Yarrowia lipolytica for the expression of human cytochrome P450 CYP1A1. Yeast 21, 583-592
Abecassis, V., Aggerbeck, L., Truan, G., and Pompon, D. (2003). Computational methods for sequence mapping of large combinatorial libraries and deduced sequence signatures. Biotechniques 34, 1280-1286
Abecassis, V., Jaffrelo, L., Rickman, D., Aggerbeck, L., Herbert, C., Truan, G., and Pompon, D. (2003). Microarray-based method for combinatorial library sequence mapping and characterization. Biotechniques 34, 1272-1279
Abecassis, V., Pompon, D., and Truan, G. (2003). Producing chimeric genes by CLERY: in vitro and in vivo recombination. In Methods in Molecular Biology, F. H. Arnold, Georgiou, G., ed. (Humana Press), pp. 165-173
Abecassis, V., Truan, G., Jaffrelo, L., and Pompon, D. (2003). Sequence mapping of combinatorial libraries on macro- or micro-arrays: experimental design of DNA arrays. In Methods in Molecular Biology, F. H. Arnold, Georgiou, G., ed. (Humana Press), pp. 189-198
Pompon, D., Truan, G., and Abecassis, V. (2003). Sequence mapping of combinatorial libraries on macro- or micro-arrays: bioinformatic treatment of data. In Methods in Molecular Biology, F. H. Arnold, Georgiou, G., ed. (Humana Press), pp. 199-211
Abecassis, V., Urban, P., Truan, G., and Pompon, D. (2003). Exploration of natural and artificial sequence spaces: towards a functionnal remodelling of membrane-bound cytochrome P450s. Biocatalysis and Biotransformation in press
Duport, C., Schoepp, B., E., C., Spagnoli, R., Dumas, B., and Pompon, D. (2003). Critical role of the plasma membrane for expression of mammalian mitochondrial side chain cleavage activity in yeast. Eur J Biochem 270, 1502-1514
Szczebara, F., Chandelier, C., Villeret, C., Masurel, A., Bourot, S., Duport, C., Blanchard, S., Groisillier, A., Testet, E., Costaglioli, P., Cauet, G., Degryse, E., Balbuena, D., Winter, J., Achstetter, T., Spagnoli, R., Pompon, D., and Dumas, B. (2003). Total biosynthesis of hydrocortisone from a simple carbon source in yeast. Nat Biotechnol 21, 143-149
Pompon, D. (2002). L'évolution moléculaire dirigée des enzymes. L'actualité chimique 8, 22-26
Urban, P. (2002) Souplesse de la spécificité de substrat des cytochromes P450, HDR, Université Paris XI
Boireau W, Bombard S, Sari MA, Pompon D. Bioengineering and characterization of DNA-protein assemblies floating on supported membranes. Biotechnol Bioeng. 2002 Jan 20;77(2):225-31.
Abecassis V, Pompon D, Truan G. Design and characterization of a novel "family-shuffling" technology adapted to membrane enzyme: application to P450s involved in xenobiotic metabolism. Adv Exp Med Biol. 2001; 500:319-22.
Loeper J, De Berardinis V, Moulis C, Beaune P, Pessayre D, Pompon D. Human epoxide hydrolase is the target of germander autoantibodies on the surface of human hepatocytes: enzymatic implications. Adv Exp Med Biol. 2001;500:121-4.
Blanquet S, Marol-Bonnin S, Beyssac E, Pompon D, Renaud M, Alric M. The 'biodrug' concept: an innovative approach to therapy. Trends Biotechnol. 2001 Oct;19(10):393-400.
Kunic B, Truan G, Breskvar K, Pompon D. Functional cloning, based on azole resistance in Saccharomyces cerevisiae, and characterization of Rhizopus nigricans redox carriers that are differentially involved in the P450-dependent response to progesterone stress. Mol Genet Genomics. 2001 Jul;265(5):930-40.
Urban P, Jobert AS, Laine R, Pompon D. Cytochrome P450 (CYP) mutants and substrate-specificity alterations: segment-directed mutagenesis applied to human CYP1A1. Biochem Soc Trans. 2001 May;29(Pt 2):128-35.
Morfin R, Lafaye P, Cotillon AC, Nato F, Chmielewski V, Pompon D.7 alpha-hydroxy-dehydroepiandrosterone and immune response. Ann N Y Acad Sci. 2000;917:971-82. Review.
Blanchard B, Pompon D, Ducrocq C. Nitrosation of melatonin by nitric oxide and peroxynitrite. J Pineal Res. 2000 Oct;29(3):184-92.
Abecassis V, Pompon D, Truan G. High efficiency family shuffling based on multi-step PCR and in vivo DNA recombination in yeast: statistical and functional analysis of a combinatorial library between human cytochrome P450 1A1 and 1A2. Nucleic Acids Res. 2000 Oct 15;28(20):E88.
De Berardinis V, Moulis C, Maurice M, Beaune P, Pessayre D, Pompon D, Loeper J. Human microsomal epoxide hydrolase is the target of germander-induced autoantibodies on the surface of human hepatocytes. Mol Pharmacol. 2000 Sep;58(3):542-51.
Louerat-Oriou B, Flinois JP, Beaune PH, Pompon D. High yield purification and characterization of engineered human P450 1A2 and generation of immuno-inhibitor antibodies. Pharmacogenetics. 1999 Feb;9(1):61-70.
Louerat-Oriou B, Perret A, Pompon D. Differential redox and electron-transfer properties of purified yeast, plant and human NADPH-cytochrome P-450 reductases highly modulate cytochrome P-450 activities. Eur J Biochem. 1998 Dec 15;258(3):1040-9.
Arand M, Muller F, Mecky A, Hinz W, Urban P, Pompon D, Kellner R, Oesch F. Catalytic triad of microsomal epoxide hydrolase: replacement of Glu404 with Asp leads to a strongly increased turnover rate. Biochem J. 1999 Jan 1;337 ( Pt 1):37-43.
Perret A, Pompon D. Electron shuttle between membrane-bound cytochrome P450 3A4 and b5 rules uncoupling mechanisms. Biochemistry. 1998 Aug 18;37(33):11412-24.
Loeper J, Louerat-Oriou B, Duport C, Pompon D. Yeast expressed cytochrome P450 2D6 (CYP2D6) exposed on the external face of plasma membrane is functionally competent. Mol Pharmacol. 1998 Jul;54(1):8-13.
Loeper J, Le Berre A, Pompon D. Topology inversion of CYP2D6 in the endoplasmic reticulum is not required for plasma membrane transport. Mol Pharmacol. 1998 Mar;53(3):408-14.
Duport C, Spagnoli R, Degryse E, Pompon D. Self-sufficient biosynthesis of pregnenolone and progesterone in engineered yeast. Nat Biotechnol. 1998 Feb;16(2):186-9.
Urban P, Mignotte C, Kazmaier M, Delorme F, Pompon D. Cloning, yeast expression, and characterization of the coupling of two distantly related Arabidopsis thaliana NADPH-cytochrome P450 reductases with P450 CYP73A5. J Biol Chem. 1997 Aug 1;272(31):19176-86.
Pompon D, Gautier JC, Perret A, Truan G, Urban P. Simulation of human xenobiotic metabolism in microorganisms. Yeast a good compromise between E. coli and human cells. J Hepatol. 1997;26 Suppl 2:81-5. Review.
Pour en savoir plus :
Le GDR2828 "Nouvelles approches en évolution dirigée des protéines"
