Group of metalloproteins

Metals lie at the heart of chemical processes that are at the origin of life and they play a crucial role in the function of numerous important proteins. Their specific properties are used in catalytic processes that are often inaccessible for active sites without metals. Conversely, the metal properties may be tuned by the protein environment in ways that are difficult to mimic in synthetic compounds.

We study the catalytic mechanisms of metalloenzymes involved in the utilisation or production of gaseous compounds like H₂, CO and CO₂ (e.g. hydrogenases, CO dehydrogenase, pyruvate:ferredoxin oxidoreductase), the insertion of metal sites in proteins and their maturation (e.g. in hydrogenases), metal transport pathways (e.g. the nickel-transporter NikA of Escherichia coli) and the regulation of metal concentrations in the cell (e.g. human iron regulatory protein 1 / cytosolic aconitase). Other aspects of our activity consist in the study of radical mechanisms of reactions catalyzed by enzymes with an S-adenosylmethionine cofactor, linked to a [4Fe-4S] cluster, and in the design of artificial metalloenzymes for the enantioselective catalysis of oxidation reactions.

The majority of the investigated proteins are very sensitive to oxygen, necessitating their study under anaerobic conditions.

Supporting programs: Biohydrogen (DSV-CEA), ANR DIVHYDO, ANR CatH2, ANR CAFE, ANR HYLIOX, ANR ENZHYd, ANR AdoMEt & ANR HAMAC.

Used methodologies: purification, crystallization and crystallokinetics of proteins under anaerobic conditions, crystallography, computational chemistry.

FRONT PAGES:

Ni-Fe hydrogenase:

(see also A Volbeda, MH Charon, C Piras, EC Hatchikian, M Frey, JC Fontecilla-Camps: "Crystal Structure of the Nickel-Iron Hydrogenase from Desulfovibrio gigas." Nature (1995), 373:580-587; and A Volbeda, E Garcin, C Piras, AL de Lacey, VM Fernandez, EC Hatchikian, M Frey, JC Fontecilla-Camps: "Structure of the [NiFe] hydrogenase active site: evidence for biologically uncommon Fe ligands." J Am Chem Soc 1996, 118:12989-12996)

Ni-Fe hydrogenase active site:

(M Frey, JC Fontecilla-Camps, A Volbeda, in Handbook of Metalloproteins; A Messerschmidt, R Huber, T Poulos, K Wieghardt, Eds.; John Wiley & Sons, Ltd: Chichester 2001, 880-896)

Structure of Fe-Fe hydrogenase:

(Y Nicolet, C Piras, P Legrand, CE Hatchikian, JC Fontecilla-Camps, Structure 1999, 7, 13-23)

Composition of the group (2009)

(e-mail : name.surname ibs.fr)

PUBLICATIONS IN 2009

Dementin S, Leroux F, Cournac L, de Lacey AL, Volbeda A, Leger C, Burlat B, Martinez N, Champ S, Martin L, Sanganas O, Haumann M, Fernandez VM, Guigliarelli B, Fontecilla-Camps JC and Rousset M, "Introduction of methionines in the gas channel makes [NiFe] hydrogenase aero-tolerant", Journal of the American Chemical Society (2009) 131:10156-10164

Fontecilla Camps JC, "Structure and Function of [NiFe]-hydrogenases", Metal Ions in Life Sciences (2009) 6:151-178

Fontecilla-Camps JC, Amara P, Cavazza C, Nicolet Y and Volbeda A, "Structure-function relationships of anaerobic gas-processing metalloenzymes", Nature (2009) 460:814-822

Goldet G, Brandmayr C, Stripp S, Happe T, Cavazza C, Fontecilla Camps JC and Armstrong FA, "Electrochemical kinetic investigations of the reactions of [FeFe]-hydrogenases with carbon monoxide and oxygen : comparing the importance of gas tunnels and active-site electronic/redox effects", Journal of the American Chemical Society (2009) 131:14979–14989

Nicolet Y, Amara P, Mouesca JM and Fontecilla-Camps JC, "Unexpected electron transfer mechanism upon AdoMet cleavage in radical SAM proteins", Proceeding of the National Academy of Sciences USA (2009) 106: 14867-14871

Pilet E, Nicolet Y, Mathevon C, Douki T, Fontecilla-Camps JC and Fontecave M, "The role of the maturase HydG in [FeFe]-hydrogenase active site synthesis and assembly", FEBS Letters (2009) 583:506-511

Reisner EF, Fontecilla-Camps JC and Armstrong FA, "Catalytic electrochemistry of a [NiFeSe]-hydrogenase on TiO2 and demostration of its suitability for visible-light driven H2 production", Chemical Communications (2009) 5:550-552

Rousselot-Pailley P, Bochot C, Marchi-Delapierre C, Jorge-Robin A, Martin L, Fontecilla-Camps JC, Cavazza C and Menage S, "The protein environment drives selectivity for sulfide oxidation by an artificial metalloenzyme", Chembiochem (2009) 10:545-552

Volbeda A, Darnault C, Tan X, Lindahl PA and Fontecilla-Camps JC, "Novel domain arrangement in the crystal structure of a truncated acetyl-CoA synthase from Moorella thermoacetica", Biochemistry (2009) 48:7916-7926

PUBLICATIONS IN 2008

Armstrong FA and Fontecilla-Camps JC, "Biochemistry. A natural choice for activating hydrogen", Science (2008) 321:498-499.

Carletti E, Li H, Li B, Ekstrom F, Nicolet Y, Loiodice M, Gillon E, Froment MT, Lockridge O, Schopfer LM, Masson P and Nachon F, "Aging of cholinesterases phosphylated by tabun proceeds through o-dealkylation", Journal of the American Chemical Society (2008) 130:16011-16020

Cavazza C, Martin L, Mondy S, Gaillard J, Ratet P and Fontecilla-Camps JC, "The possible role of an [FeFe]-hydrogenase-like protein in the plant responses to changing atmospheric oxygen levels", Journal of Inorganic Biochemistry (2008) 102:1359-1365.

Cherrier MV, Cavazza C, Bochot C, Lemaire D and Fontecilla-Camps JC, "Structural characterization of a putative endogenous metal chelator in the periplasmic nickel transporter NikA", Biochemistry (2008) 47:9937-9943.

Fdez Galván I, Volbeda A, Fontecilla-Camps JC and Field MJ, "A QM/MM study of proton transport pathways in a [NiFe] hydrogenase", Proteins: Structure, Function and Bioinformatics (2008), 73:195-203

Leroux F, Dementin S, Burlat B, Cournac L, Volbeda A, Champ S, Martin L, Guigliarelli B, Bertrand P, Fontecilla-Camps J, Rousset M and Leger C, "Experimental approaches to kinetics of gas diffusion in hydrogenase", Proceedings of the National Academy of Sciences U S A (2008) 105: 11188-11193

Nicolet Y, Rubach JK, Posewitz MC, Amara P, Mathevon C, Atta M, Fontecave M, Fontecilla-Camps JC, "X-ray structure of the [FeFe]-hydrogenase maturase hydE from Thermotoga maritima", Journal of Biological Chemistry (2008), 283:18861-18872

Parkin A, Goldet G, Cavazza C, Fontecilla-Camps JC and Armstrong FA, " The difference a Se makes? Oxygen-Tolerant hydrogen production by the [NiFeSe]-hydrogenase from Desulfomicrobium baculatum", Journal of the American Chemical Society (2008) 130:13410-13416

PUBLICATIONS IN 2007

Amara P, Fdez Galvan I, Fontecilla-Camps JC and Field MJ, "The enamine intermediate may not be universal to thiamine catalysis", Angewandte Chemie International Edition England (2007) 46: 9019-9022

Chandor A, Douki T, Gasparutto D, Gambarelli S, Sanakis Y, Nicolet Y, Ollagnier-de-Choudens S, Atta M and Fontecave M, "Characterization of the DNA repair spore photoproduct lyase enzyme from Clostridium acetobutylicum: A radical-SAM enzyme", Comptes Rendus Chimie (2007) 10(8): 756-765

Fontecilla-Camps JC, Volbeda A, Cavazza C and Nicolet Y, "Structure/function relationships of [NiFe]- and [FeFe]-hydrogenases", Chemical Reviews (2007) 107: 4273-4303

Morales R, Berna A, Carpentier P, Contreras-Martel C, Renault F, Nicodeme M, Chesne-Seck ML, Bernier F, Dupuy J, Schaeffer C, Diemer H, Van-Dorsselaer A, Fontecilla-Camps JC, Masson P, Rochu D and Chabriere E, "DŽcouverte et structure cristallographique dÕune apolipoprotŽine humaine", Annales Pharmaceutiques Françaises (2007) 65(2): 98-107

Razavet M, Artero V, Cavazza C, Oudart Y, Lebrun C, Fontecilla-Camps JC and Fontecave M,"Tricarbonylmanganese(I)-lysosyme complex: A structurally characterized organometallic protein", Chemical Communications (2007) 27: 2805-2807

PUBLICATIONS IN 2006

Cavazza C, Contreras-Martel C, Pieulle L, Chabriere E, Hatchikian EC and Fontecilla-Camps JC. Flexibility of thiamine diphosphate revealed by kinetic crystallographic studies of the reaction of pyruvate-ferredoxin oxidoreductase with pyruvate. Structure (2006) 14: 217-224.

Contreras-Martel C, Carpentier P, Morales R, Renault F, Chesne-Seck ML, Rochu D, Masson P, Fontecilla-Camps JC and Chabriere E, "Crystallization and preliminary X-ray diffraction analysis of human phosphate-binding protein", Acta Crystallographica Section F: Structural Biology and Crystallization Communications (2006) 62: 67-69

Dupuy J, Volbeda A, Carpentier P, Darnault C, Moulis JM and Fontecilla-Camps JC. Crystal structure of human iron regulatory protein 1 as cytosolic aconitase. Structure (2006) 14: 129-139

Morales R, Berna A, Carpentier P, Contreras-Martel C, Renault F, Nicodeme M, Chesne-Seck ML, Bernier F, Dupuy J, Schaeffer C, Diemer H, Van-Dorsselaer A, Fontecilla-Camps JC, Masson P, Rochu D and Chabriere E, "Serendipitous discovery and x-ray structure of a human phosphate binding apolipoprotein", Structure (2006) 14: 601-609

Parkin A, Cavazza C, Fontecilla-Camps JC and Armstrong FA, "Electrochemical investigations of the interconversions between catalytic and inhibited states of the [FeFe]-hydrogenase from Desulfovibrio desulfuricans", Journal of the American Chemical Society (2006) 128: 16808-16815

Pecqueur L, D’Autréaux B, Dupuy J, Nicolet Y, Jacquamet L, Brutscher B, Michaud-Soret I and Bersch B. Structural changes of Escherichia coli ferric uptake regulator during metal-dependent dimerization and activation explored by NMR and x-ray crystallography. Journal of Biological Chemistry (2006) 281: 21286-21295

Tan X, Volbeda A, Fontecilla-Camps J and Lindahl P. Function of the tunnel in acetylcoenzyme A synthase / carbon monoxide dehydrogenase. Journal of Biological Inorganic Chemistry (2006) 11: 371-378

Volbeda A and Fontecilla-Camps JC. "Catalytic nickel-iron sulfur clusters: from minerals to enzymes." Topics in Organometallic Chemistry (2006) 17, 57-82

Walden WE, Selezneva AI, Dupuy J, Volbeda A, Fontecilla-Camps JC, Theil EC and Volz K. Crystal structure of iron regulatory protein 1 complexed with ferritin IRE-RNA. Science (2006) 314, 1903-1908.

PUBLICATIONS IN 2005

Amara P, Volbeda A, Fontecilla-Camps JC and Field MJ. A quantum chemical study of the reaction mechanism of acetyl-coenzyme a synthase. Journal of the American Chemical Society (2005) 127: 2776-2784

Cherrier M, Martin L, Cavazza C, Jacquamet L, Lemaire D, Gaillard J and Fontecilla-Camps JC. Crystallographic and spectroscopic evidence for high affinity binding of FeEDTA(H2O)- to periplasmic nickel transporter NikA. Journal of the American Chemical Society (2005) 127: 10075-10082

Dupuy J, Darnault C, Brazzolotto X, Kuhn LC, Moulis JM, Volbeda A and Fontecilla-Camps JC. Crystallization and preliminary X-ray diffraction data for the aconitase form of human iron-regulatory protein 1. Acta Crystallographica Section F Structural Biology and Crystallization Communications (2005) 61: 482-485

Tormos JR, Wiley KL, Seravalli J, Nachon F, Masson P, Nicolet Y, Quinn DM. The reactant state for substrate-activated turnover of acetylthiocholine by butyrylcholinesterase is a tetrahedral intermediate. Journal of the American Chemical Society (2005) 127:14538-14539.

Vincent KA, Parkin A, Lenz O, Albracht SP, Fontecilla-Camps JC, Cammack R, Friedrich B and Armstrong FA. Electrochemical definitions of O2 sensitivity and oxidative inactivation in hydrogenases. Journal of the American Chemical Society (2005) 127: 18179-18189

Volbeda A and Fontecilla-Camps JC. Structure-function relationships of nickel-iron sites in hydrogenase and a comparison with the active sites of other nickel-iron enzymes. Coordination Chemistry Reviews (2005) 249: 1609-1619

Volbeda A and Fontecilla-Camps JC. Structural bases for the catalytic mechanism of Ni-containing carbon monoxide dehydrogenases. Dalton Transactions (2005) 3443-3450

Volbeda A, Martin L, Cavazza C, Matho M, Faber BW, Roseboom W, Albracht SPJ, Garcin E, Rousset M and Fontecilla-Camps JC. Structural differences between the ready and unready oxidized states of [NiFe] hydrogenases. Journal of Biological Inorganic Chemistry (2005) 10: 239-249

PUBLICATIONS IN 2000-2004

Chabriere E, Vernede X, Guigliarelli B, Charon MH, Hatchikian EC and Fontecilla-Camps JC. Crystal structure of the free radical intermediate of pyruvate:ferredoxin oxidoreductase. Science (2001) 294: 2559-63.

Contreras Martel C, Martinez Oyanedel J, Bunster M, Legrand P, Piras C, Vernede X and Fontecilla Camps JC. Crystallization and 2.2 angstrom Å resolution structure of R-phycoerythrin from Gracilaria chilensis: a case of perfect hemihedral twinning. Acta Crystallographica Section D Biological Crystallography (2001) 57: 52-60

Darnault C, Volbeda A, Kim EJ, Legrand P, Vernede X, Lindahl PA and Fontecilla-Camps JC. Ni-Zn-[Fe(4)-S(4)] and Ni-Ni-[Fe(4)-S(4)] clusters in closed and open alpha subunits of acetyl-CoA synthase/carbon monoxide dehydrogenase. Nature Structural Biology (2003) 10: 271-279

Frey M. Hydrogenases: Hydrogen-activating enzymes. Chembiochem (2002) 3: 153-160

Hurley JK, Morales R, Martinez Julvez M, Brodie TB, Medina M, Gomez Moreno C and Tollin G. Structure-function relationships in Anabaena ferredoxin/ferredoxin: NADP(+) reductase electron transfer: insights from site-directed mutagenesis, transient absorption spectroscopy and X-ray crystallography. Biochimica Et Biophysica Acta Bioenergetics (2002) 1554: 5-21

Loudwig S, Nicolet Y, Masson P, Fontecilla-Camps JC, Bon S, Nachon F, Goeldner M. Photoreversible inhibition of cholinesterases: catalytic serine-labeled caged butyrylcholinesterase. Chembiochem (2003) 4: 762-767.

Morales R, Charon MH, Hudry-Clergeon G, Petillot Y, Norager S, Medina M and Frey M. Refined X-ray Structures of the Oxidized, at 1.3 A, and Reduced, at 1.17 A, [2Fe-2S] Ferredoxin from the Cyanobacterium Anabaena PCC7119 Show Redox-Linked Conformational Changes. Biochemistry (2000) 39: 2428.

Morales R, Charon MH, Kachalova G, Serre L, Medina M, Gomez Moreno C and Frey M. A redox-dependent interaction between two electron-transfer partners involved in photosynthesis. EMBO Reports (2000) 1: 271-276

Morales R, Frey M and Mouesca JM. An approach based on quantum chemistry calculations and structural analysis of a [2Fe-2S*] ferredoxin that reveal a redox-linked switch in the electron-transfer process to the Fd-NADP(+) reductase. Journal of the American Chemical Society (2002) 124: 6714-6722

Morales R, Kachalova G, Vellieux F, Charon MH and Frey M. Crystallographic studies of the interaction between the ferredoxin- NADP+ reductase and ferredoxin from the cyanobacterium Anabaena: looking for the elusive ferredoxin molecule. Acta Crystallographica D Biological Crystallography (2000) 56: 1408-1412

Morelli X, Czjzek M, Hatchikian CE, Bornet O, Fontecilla-Camps JC, Palma PN, Moura JJ and Guerlesquin F, Structural model of the Fe-hydrogenase-cytochrome c553 complex combining TROSY experiments and soft docking calculations. Journal of Biological Chemistry (2000) 275: 23204-10

Nachon F, Nicolet Y, Viguie N, Masson P, Fontecilla Camps JC and Lockridge O. Engineering of a monomeric and low-glycosylated form of human butyrylcholinesterase - Expression, purification, characterization and crystallization. European Journal of Biochemistry (2002) 269: 630-637

Nicolet Y, Cavazza C and Fontecilla-Camps JC. Fe-only hydrogenases: structure, function and evolution. Journal of Inorganic Biochemistry (2002) 91: 1-8.

Nicolet Y, De Lacey AL, Vernede X, Fernandez VM, Hatchikian EC and Fontecilla Camps JC. Crystallographic and FTIR spectroscopic evidence of changes in Fe coordination upon reduction of the active site of the Fe-only hydrogenase from Desulfovibrio desulfuricans. Journal of the American Chemical Society (2001) 123: 1596-1601

Nicolet I, Lemon BJ, Fontecilla-Camps JC and Peters JW. A novel FeS cluster in Fe-only hydrogenases. Trends in Biochemical Sciences (2000) 25: 138-43.

Nicolet Y, Lockridge O, Masson P, Fontecilla-Camps JC and Nachon F. Crystal structure of human butyrylcholinesterase and of its complexes with substrate and products. The Journal Of Biological Chemistry (2003) 278: 41141-41147

Stadler C, De Lacey AL, Montet Y, Volbeda A, Fontecilla-Camps JC, Conesa JC and Fernandez VM. Density Functional Calculations for Modeling the Active Site of Nickel- Iron Hydrogenases. 2. Predictions for the Unready and Ready States and the Corresponding Activation Processes. Inorganic Chemistry (2002) 41: 4424-4434

Volbeda A and Fontecilla-Camps JC. Crystallographic evidence for a CO/CO(2) tunnel gating mechanism in the bifunctional carbon monoxide dehydrogenase/acetyl coenzyme A synthase from Moorella thermoacetica. Journal Of Biological Inorganic Chemistry (2004) 9: 525-532

Volbeda A and Fontecilla Camps JC. The active site and catalytic mechanism of [NiFe] hydrogenases. Dalton Transactions (2003) 4030-4038