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Structure and function of quinone binding sites

par grimaldi - publié le , mis à jour le

Our goal is to elucidate the molecular factors responsible for reactivity and specificity of quinone utilization by membrane-bound bioenergetic complexes that couple electron transfer to the generation of a transmembrane proton electrochemical gradient during bacterial respiration.
Our efforts are mainly directed at the membrane-bound nitrate reductase A (NarGHI) from Escherichia coli, an energy transducing complex optimally induced in anaerobiosis and in high nitrate concentration in the growth medium. This heterotrimeric metalloenzyme is able to oxidize ubiquinols (primarily involved in aerobic respiration), menaquinols (electron carriers in anaerobic respiration) and demethylmenaquinols, and to stabilize their paramagnetic semiquinone intermediate at a single site. Thus, it constitutes an ideal model system to address at the molecular level the question of the adaptation of an anaerobic enzyme to oxygenic conditions.

We use multifrequency high-resolution Electron Paramagnetic Resonance (EPR) spectroscopy to provide detailed structural information at atomic resolution on the semiquinone binding mode via the analysis of weak magnetic and quadrupole interactions from nearby paramagnetic nuclei (e.g. 1H, 2H, 14N, 15N), These can be detected using Electron Nuclear Double Resonance (ENDOR) spectroscopy or multidimensional pulsed EPR techniques based on the measurement of electron spin echoes (Electron spin echo envelope modulation – ESEEM – Hyperfine Sublevel Correlation spectroscopy – HYSCORE, Figure 2). The use of a multifrequency approach greatly increases the power of these methods.

Publications :

  • M. Seif Eddine, F. Biaso, R. Arias-Cartin, E. Pilet, J. Rendon, S. Lyubenova, F. Seduk, B. Guigliarelli, A. Magalon & S. Grimaldi*
    Probing the menasemiquinone binding mode to nitrate reductase A by selective 2H & 15N labelling, HYSCORE spectroscopy and DFT modeling
    ChemPhysChem (2017), 18, 2704-2714
  • J. Rendon, E. Pilet, Z. Fahs, F. Seduk, L. Sylvi, M. Hajj Chehade, F. Pierrel, B. Guigliarelli, A. Magalon* & S. Grimaldi*
    Demethylmenaquinol is a substrate of Escherichia coli nitrate reductase A (NarGHI) and forms a stable semiquinone intermediate at the NarGHI quinol oxidation site
    Biochim. Biophys. Acta - Bioenergetics (2015), 1847, 739-747
  • S. Grimaldi, B. Schoepp-Cothenet, P. Ceccaldi, B. Guigliarelli & A. Magalon*
    The prokaryotic Mo/W-bisPGD enzymes family : a catalytic workhorse in bioenergetics
    Biochim. Biophys. Acta - Bioenergetics (2013),1827, 1048-1085
  • R. Arias-Cartin, S. Grimaldi, P. Arnoux, B. Guigliarelli & A. Magalon*
    Cardiolipin binding in bacterial respiratory complexes : Structural and functional implications
    Biochim. Biophys. Acta - Bioenergetics (2012), 1817, 1937-1949
  • S. Grimaldi*, R. Arias-Cartin, P. Lanciano, S. Lyubenova, R. Szenes, B. Endeward, T. F. Prisner, B. Guigliarelli & A. Magalon
    Determination of the proton environment of high stability menasemiquinone intermediate in Escherichia coli nitrate reductase A by pulsed EPR
    J. Biol. Chem. (2012), 287, 4662-4670
  • R. Arias-Cartin, S. Grimaldi, J. Pommier, P. Lanciano, C. Schaefer, P. Arnoux, G. Giordano, B. Guigliarelli & A. Magalon*
    Cardiolipin-based respiratory complex activation in bacteria
    Proc. Nat. Acad. Sci. USA (2011), 108, 7781-7786
  • F. MacMillan*, S. Kacprzak, P. Hellwig, S. Grimaldi, H. Michel, & M. Kaupp
    Elucidating mechanisms in haem copper oxidases : The high-affinity QH binding site in quinol oxidase as studied by DONUT-HYSCORE spectroscopy and density functional theory
    Faraday Discussions (2011), 148, 315-344
  • R. Arias-Cartin, S. Lyubenova, P. Ceccaldi, T. F. Prisner, A. Magalon, B. Guigliarelli & S. Grimaldi*
    HYSCORE evidence that mena- and ubisemiquinones bind at the same Q-site (QD) of E. coli nitrate reductase A
    J. Am. Chem. Soc. (2010), 132, 5942-5943
  • S. Grimaldi*, R. Arias-Cartin, P. Lanciano, S. Lyubenova, B. Endeward, T. F. Prisner, A. Magalon & B. Guigliarelli
    Direct evidence for nitrogen ligation to the high-stability semiquinone intermediate in E. coli nitrate reductase A
    J. Biol. Chem. (2010), 285, 179-187
  • P. Lanciano, A. Magalon, P. Bertrand, B. Guigliarelli & S. Grimaldi*
    High-stability semiquinone intermediate in the nitrate reductase A (NarGHI) from Escherichia coli is located in a quinol oxidation site close to heme bD
    Biochemistry (2007), 46, 5323-5329
  • S. Grimaldi*, P. Lanciano, P. Bertrand, F. Blasco, & B. Guigliarelli
    Evidence for an EPR-detectable semiquinone intermediate in the membrane-bound subunit NarI of nitrate reductase A (NarGHI) from Escherichia coli
    Biochemistry (2005), 44, 1300-1308

Contact : grimaldi"at"