Florence Lederer
DR émérite, CNRS
Laboratoire de Chimie Physique
Université Paris-Sud, 91405 Orsay Cedex
Tel : (33) 1 69 15 56 12
Fax : (33) 1 69 15 61 88

Research interests
Enzymes chemical mechanisms, in particular flavoenzymes. Protein structure and protein chemistry. NADPH oxidase from neutrophils (Nox 2), assembly and disassembly.

Methods
Kinetic and spectrophotometric studies of recombinant wild-type and mutant enzymes, using substrates, substrates analogues, inhibitors, inactivators, suicide reagents. Study of protein-protein interactions via Surface plasmon resonance (Biacore) and using monoclonal antibodies. Manipulation of neutrophils from human blood.

Collaborations with crystallographers, theoretical chemists, kineticists, in France, Europe, the USA and Japan.

Results
Definition of the family of b5-like cytochromes and of L-2-hydroxy acid-oxidizing flavoenzymes. Characterization of the chemical mechanism of these flavoenzymes as involving a carbanion. Definition of the interface between the two domains of flavocytochrome b2 which is competent for electron transfer. Characterization of the epitope of a monoclonal antibody that disrupts the functional contacts between these two domains.
Demonstration by affinity labelling that gp91, one of the Nox2 membrane subunits, carries the NADPH binding site. Inhibition of Nox2 activation by a reagent that prevents phosphorylation of p47phox and complex assembly, and also inhibits activity by dephosphorylation and disassembly.

Career
PhD in Physical Sciences, Strasbourg University, 1963
Two post doctoral years in the Department of Chemistry, Harvard University (1963-65).
CNRS researcher from 1960 to 2003, now director of research emeritus. Work at the Centre de Génétique Moléculaire, CNRS, Gif-sur-Yvette (1965-1984), INSERM U70, Hôpital Necker, Paris (1984-1995), Laboratoire d'Enzymologie et Biochimie Structurales, CNRS, Gif-sur-Yvette (1996-2007).  Now  at Laboratoire de Chimie Physique, Faculté des Sciences, Orsay.

Recent publications

  1. Kinetic and crystallographic studies on the active-site Arg289Lys mutant of flavocytochrome b2 (yeast L-lactate dehydrogenase). C. G. Mowatt, I. Beaudoin, R. C. E. Durley, J. D. Barton, A. D. Pike, Chen, Z.-w, G. A. Reid, S. K. Chapman, F. S. Mathews and F. Lederer (2000) Biochemistry, 39, 3266-3275.
  2. Flavin-protein interactions in flavocytochrome b2 as studied by NMR after reconstitution of the enzyme with 13C- and 15N-labelled flavin. G. Fleischmann, F. Lederer, F. Müller, A. Bacher and H. Rüterjans (2000) Eur. J. Biochem., 267, 5156-5167.
  3. The catalytic role of tyrosine 254 in flavocytochrome b2  (L-lactate dehydrogenase from baker's yeast). Comparison between the Y254F and Y254L mutant proteins. M. Gondry, J. Dubois, M. Terrier and F. Lederer (2001) Eur. J. Biochem., 268, 4918- 4927.
  4. Hydroxamates as substrates and inhibitors for FMN-dependent 2-hydroxy acid dehydrogenases. D. Amar, P. North, V.Miskiniene, N. Cénas & F. Lederer (2002) Bioorg. Chem. 30, 145-162.
  5. Epitope mapping for the monoclonal antibody that inhibits intramolecular electron transfer in flavocytochrome b2. K. H. D. Lê, M. Mayer and F. Lederer (2003) Biochem. J. 373, 115-123.
  6. Interaction of quinones with thioredoxin reductase. A challenge to the antioxidant role  of the mammalian selenoprotein. N. Cénas, H. Nivinskas, Z. Anusevicius, J. Sarlauskas, F. Lederer and E. S. J. Arnér (2004) J. Biol. Chem., 279, 2583-2592.
  7. Another biological effect  of tosylphenylalaninechloromethane (TPCK): it prevents p47phox phosphorylation and translocation upon neutrophil stimulation. M. Gillibert, Z. Dehry, M. Terrier, J. El Benna and F. Lederer (2005) Biochem. J.  386, 549-556.
  8. Crystal structure analysis of recombinant rat kidney long-chain hydroxy acid oxidase L. M. Cunane, J. D. Barton, Z.-w. Chen, K. H. D. Lê, D. Amar, F. Lederer and F. S.   Mathews, (2005) Biochemistry, 44, 1521-1531.
  9. Which mechanism for (S)-2-hydroxy acid-oxidizing flavoenzymes? A tale of a histidine and two protons. F. Lederer, D.Amar, A. K. Ould Boubacar and C.Vignaud (2005) in Flavins and Flavoproteins 2005, Nishino, T, Miura, R., Tanokura, M and Fukui, K. edts, ArchiTect Inc. Tokyo, pp 193-204.
  10. Cyclopropylglycolate, a new substrate and inactivator for long-chain hydroxy acid oxidase and flavocytochrome b2. A. K. Ould Boubacar, K.K. Frederick, V. Miskiniene,  N. Cénas and F. Lederer in Flavins and Flavoproteins 2005, Nishino, T, Miura, R., Tanokura, M and Fukui, K. edts, ArchiTect Inc. Tokyo, pp 419-423.
  11. A kinetic distinction between the flavin reduction step and the first electron transfer to the heme for two flavocytochrome b2 mutants. K. H. D. Lê, A.Boussac and F. Lederer (2005) in Flavins and Flavoproteins 2005, Nishino, T, Miura, R., Tanokura, M and Fukui, K. edts, ArchiTect Inc. Tokyo, pp 443-448.
  12. Ab initio molecular dynamics studies of flavocytochrome b2 reduction. G. Tabacchi, M. A. Vanoni, F. Lederer, A. Gamba and E. Fois (2005) in Flavins and Flavoproteins 2005, Nishino, T, Miura, R., Tanokura, M and Fukui, K. edts, ArchiTect Inc. Tokyo, pp 281-286.
  13. Electron flow in multicenter enzymes : theory, applications and consequences on the natural design of redox chains. C. Léger, F. Lederer, B. Guigliarelli and P. Bertrand (2006) J. Am. Chem. Soc.,128, 180-187.
  14. Molecular characterization of laforin, a dual-specificity protein phosphatase implicated in Lafora disease. J. M. Girard, K. H. D. Lê and F. Lederer (2006) Biochimie, 88, 1961-1971.
  15. Potentiometric and further kinetic characterization of the flavin-binding domain of Saccharomyces cerevisiae  flavocytochrome b2. Inhibition by anions binding in the active site. N. Čénas, K.H. D. Lê, M. Terrier and F. Lederer (2007) Biochemistry, 46, 4661-4670.
  16. Purification and characterization of recombinant human liver glycolate oxidase. C. Vignaud,  N. Pietrancosta, E. L.Williams, G. Rumsby and F. Lederer (2007), Arch.  Biochem. Biophys. 465, 410-416.
  17. Flavocytochrome b2 : Reactivity of its flavin with molecular oxygen. A. K. O. Boubacar,  S. Pethe, J. P. Mahy and F. Lederer (2007) Biochemistry, 46, 13080- 13088.
  18. Mobility and electron transfer in flavocytochrome b2 . K. H. D. Lê, B. Golinelli-Pimpaneau and F. Lederer (2008) in Flavins and Flavoproteins 2008, S. Frago, C. Gómez-Moreno and M. Medina, Edts, Prensas Universitarias de Zaragoza, pp147-156.
  19. L-lactate dehydrogenation in flavocytochrome b2: A first principles molecular dynamics study.G. Tabacchi, D.Zucchini, G. Caprini, A. Gamba, F. Lederer,|M.A. Vanoni and E.Fois (2009) FEBS J., 276, 2368-2380.
  20. Crystal structure of human glycolate oxidase in complex with the inhibitor 4-carboxy-5- [(4-chlorophenyl)sulfanyl]-1, 2, 3-thiadiazole, J.M.Bourhis, C.Vignaud, N.Pietrancosta, F. Guéritte, D. Guénard, F.Lederer and Y.Lindqvist (2009) Acta Cryst., F65, 1246-1253.
  21. Interdomain contacts in flavocytochrome b2 , a mutational analysis K. H. D. Lê, A. Boussac, B.Frangioni, C. Léger and F. Lederer (2009) Biochemistry, 48, 10803-10809.