Pierre LAFITE

LAFITE Pierre

Assistant Professor, HDR, University of Orléans

Enzymology and glycobiochemistry
Institut de Chimie Organique et Analytique
université d'Orléans - Pôle de chimie
rue de Chartres - BP 6759
45067 Orléans Cedex 2
France

pierre.lafite@univ-orleans.fr
Phone: 02 38 49 48 55



Research themes: 

I mainly focus on the development and the use of enzyme to generate glycosides. This objective is supported by the use of many techniques, ranging from molecular biology (cloning, protein engineering), biochemistry (protein characterization, enzymology), to biocatalysis and structural biology (molecular modeling, crystallography).

The targeted compounds to synthesize are expected to exhibit innovative properties in pharmaceutics, biochemistry or cosmetics.

 

Biography: 

Having initially obtained a BSc and an master degree in Physical Chemistry in 2001 and 2002 in Ecole Normale supérieure in Lyons, I shifted on Molecular Enzymology during my 2nd year Master degree and my PhD at UMR8601 (Paris), under the supervision of Dr D. Mansuy. I then spend 2 yers as a Post-Doctoral Research Associate at the Manchester Institute of Biotechnology, in Profs. Scrutton and Leys group. I was finally recruited in 2009 as an assistant professor of biochemistry in ICOA laboratory, and received my Habilitation to conduct research in 2019.

Administrative responsibilities: 

Teaching :

Life Sciences BSc: General biochemistry, Experimental enzymology, Structural biology and protein purification

Life Sciences Master degree: Protein crystallography, protein engineering

Molecular chemistry Master degree: general biochemistry.

Bibliographic Links and Social Networks: 

   

 

Publications: 

Peer-reviewed Publications:

37- Guillotin, L., Assaf, Z., Pistorio, S.G., Lafite, P., Demchenko, A. V., Daniellou, R. Hydrolysis of Glycosyl Thioimidates by Glycoside Hydrolase Requires Remote Activation for Efficient Activity. (2019) Catalysts 9, 826. https://doi.org/10.3390/catal9100826

36- Lafite P.*, Marroun S., Coadou G., Montaut S., Marquès S., Schuler M., Rollin P., Tatibouët A., Daniellou R., Oulyadi H. (2019) S-Glycosyltransferase UGT74B1 can glycosylate both S- and O-acceptors: mechanistic insights through substrate specificity. Mol. Catal. 479, 110631 https://doi.org/10.1016/j.mcat.2019.110631

35- Seničar M., Legentil L., Ferrières V., Eliseeva S.V., Petoud S., Takegawa K., Lafite P., Daniellou R. (2019). Galactofuranosidase from JHA 19 Streptomyces sp.: subcloning and biochemical characterization. Carbohyd. Res., 480, 35-41. https://www.doi.org/10.1016/j.carres.2019.05.011

34- Ferey J., Da Silva D., Colas C., Nehmé R., Lafite P., Roy V., Morin P., Daniellou R., Agrofoglio L., Maunit B. (2019). Monitoring of successive phosphorylations of thymidine using free and immobilized human nucleoside/nucleotide kinases by Flow Injection Analysis with High-Resolution Mass Spectrometry. Anal. Chim. Acta, 1049, 115-122. https://doi.org/10.1016/j.aca.2018.10.032

33- Guillotin, L., Kim, H., Traore, Y., Moreau, P., Lafite, P., Coquoin, V., Nuccio, S., de Vaumas, R. & Daniellou, R. (2019). Biochemical Characterization of the α-L-Rhamnosidase DtRha from Dictyoglomus thermophilum : Application to the Selective Derhamnosylation of Natural Flavonoids ACS Omega. 4, 1916–1922. https://www.doi.org/10.1021/acsomega.8b03186

32- Ati, J., Colas, C., Lafite, P., Sweeney, R. P., Zheng, R. B., Lowary T. L., Daniellou, R. (2018) The LPG1x family from Leishmania major is constituted of rare eukaryotic galactofuranosyltransferases with unprecedented catalytic properties. Sci. Rep., 8(1), 17566. https://www.doi.org/10.1038/s41598-018-35847-w

31- Ferey, J., Da Silva, D., Colas, C., Nehmé, R., Lafite, P., Roy, V., Morin, P., Daniellou, R., Agrofoglio, L., Maunit, B. (2019) Monitoring of successive phosphorylations of thymidine using free and immobilized human nucleoside/nucleotide kinases by Flow Injection Analysis with High-Resolution Mass Spectrometry. Anal. Chim. Acta, 1049, 115–122.https://www.doi.org/10.1016/j.aca.2018.10.032

30- Lafite, P., André, F., Graves, J., Zeldin, D., Dansette, P., Mansuy, D. (2018) Role of Arginine 117 in Substrate Recognition by Human Cytochrome P450 2J2. Int. J. Mol. Sci. 19, 2066. https://www.doi.org/10.3390/ijms19072066

29- Guillotin, L., Richet, N., Lafite, P.*, Daniellou, R. (2017). Is the acid/base catalytic residue mutation in β-D-mannosidase DtMan from Dictyoglomus thermophilum sufficient to provide thioglycoligase activity? Biochimie, 137, 190–196. https://www.doi.org/10.1016/j.biochi.2017.03.020

28- Guillotin, L., Cancellieri, P., Lafite, P., Landemarre, L., Daniellou, R., Corzo, N., Daniellou, R. (2017). Chemo-enzymatic synthesis of 3-O-(β-D-glycopyranosyl)-sn-glycerols and their evaluation as preservative in cosmetics. Pure App. Chem., 89(9), 11302. https://www.doi.org/10.1515/pac-2016-1210

27- Ferey, J., Da Silva, D., Lafite, P., Daniellou, R., Maunit, B. (2017). TLC-UV hyphenated with MALDI-TOFMS for the screening of invertase substrates in plant extracts. Talanta, 170, 419–424.https://www.doi.org/10.1016/j.talanta.2017.04.040

26- Ortmayer, M., Lafite, P., Menon, B. R. K., Tralau, T., Fisher, K., Denkhaus, L., Leys, D. (2016). An oxidative N-demethylase reveals PAS transition from ubiquitous sensor to enzyme. Nature, 539(7630), 593–597. https://www.doi.org/10.1038/nature20159

25- Marroun, S., Montaut, S., Marquès, S., Lafite, P., Coadou, G., Rollin, P., Jousset, G., Tatibouët A, Oulyadi, H, Daniellou, R. (2016). UGT74B1 from Arabidopsis thaliana as a versatile biocatalyst for the synthesis of desulfoglycosinolates. Org. Biomol. Chem., 14(26), 6252–6261. https://www.doi.org/10.1039/c6ob01003b

24- Ferey, J., Da Silva, D., Bravo-Veyrat, S., Lafite, P., Daniellou, R., Maunit, B. (2016). Validation of a thin-layer chromatography/densitometry method for the characterization of invertase activity. J. Chrom. A, 1477, 108-113. https://www.doi.org/ 10.1016/j.chroma.2016.11.049

23- Sauvageot, E., Lafite, P., Duverger, E., Marion, R., Hamel, M., Gaillard, S., Daniellou, R. (2016). Iridium complexes inhibit tumor necrosis factor-alpha by utilizing light and mixed ligands. J. Organomet. Chem., 808, 122–127. https://www.doi.org/ 10.1016/j.jorganchem.2016.02.001

22- Belaz, S., Rattier, T., Lafite, P., Moreau, P., Routier, F. H., Robert-Gangneux, F., Gangneux, J-P., Daniellou, R. (2015). Identification, biochemical characterization, and in-vivo expression of the intracellular invertase BfrA from the pathogenic parasite Leishmania major. Carbohydr. Res., 415, 31–38. https://www.doi.org/ 10.1016/j.carres.2015.07.001

21- Fayad, S., Nehmé, R., Lafite, P., & Morin, P. (2015). Assaying human neutrophil elastase activity by capillary zone electrophoresis combined with laser-induced fluorescence. J. Chrom. A. 1419, 116-124. https://www.doi.org/ 10.1016/j.chroma.2015.09.084

20- François-Heude, M., Méndez-Ardoy, A., Cendret, V., Lafite, P., Daniellou, R., Ortiz Mellet, C., Garcìa Fernandez, J. M., Moreau, V., Djedaïni-Pilard, F. (2015). Synthesis of High-Mannose Oligosaccharide Analogues through Click Chemistry: True Functional Mimics of Their Natural Counterparts Against Lectins? Chem. Eur. J., 21(5), 1978–1991. https://www.doi.org/10.1002/chem.201405481

19- Guillotin, L., Lafite, P., Daniellou, R. (2014). Unraveling the Substrate Recognition Mechanism and Specificity of the Unusual Glycosyl Hydrolase Family 29 BT2192 from Bacteroides thetaiotaomicron. Biochemistry, 53(9), 1447–1455. https://www.doi.org/10.1021/bi400951q

18- Nehmé, R., Nehmé, H., Saurat, T., de-Tauzia, M.-L., Buron, F., Lafite, P., Verrelle, P., Chautard, E., Morin, P., Routier, S., Bénédetti, H. (2014). New in-capillary electrophoretic kinase assays to evaluate inhibitors of the PI3k/Akt/mTOR signaling pathway. Anal. Bioanal. Chem., 406(15), 3743–3754. https://www.doi.org/ 10.1007/s00216-014-7790-z

17- Amaral, M., Levy, C., Heyes, D. J., Lafite, P., Outeiro, T. F., Giorgini, F., Leys, D.,  Scrutton, N. S. (2013). Structural basis of kynurenine 3-monooxygenase inhibition. Nature, 496(7445), 382–385. https://www.doi.org/10.1038/nature12039

16- Barkauskaite, E., Brassington, A., Tan, E. S., Warwicker, J., Dunstan, M. S., Banos, B., Lafite P., Ahel I., Leys, D. (2013). Visualization of poly(ADP-ribose) bound to PARG reveals inherent balance between exo- and endo-glycohydrolase activities. Nat. Commun., 4, 2164. https://www.doi.org/10.1038/ncomms3164

15- Nehmé, H., Nehmé, R., Lafite, P., Routier, S., & Morin, P. (2013). In-capillary reactant mixing for monitoring glycerol kinase kinetics by capillary electrophoresis. J. Sep. Sci., 36(13), 2151–2157. https://www.doi.org/10.1002/jssc.201300063

14- Nehmé, H., Nehmé, R., Lafite, P., Duverger, E., Routier, S., Morin, P. (2013). Electrophoretically mediated microanalysis for in-capillary electrical cell lysis and fast enzyme quantification by capillary electrophoresis. Anal. Bioanal. Chem., 405(28), 9159–9167. https://www.doi.org/10.1007/s00216-013-7332-0

13- Nehmé, H., Nehmé, R., Lafite, P., Routier, S., Morin, P. (2013). Human protein kinase inhibitor screening by capillary electrophoresis using transverse diffusion of laminar flow profiles for reactant mixing. J. Chrom. A, 1314, 298–305. https://www.doi.org/ 10.1016/j.chroma.2013.08.046

12- Dunstan, M. S., Barkauskaite, E., Lafite, P., Knezevic, C. E., Brassington, A., Ahel, M., Hergenrother, P.J., Leys, D., Ahel, I. (2012). Structure and mechanism of a canonical poly(ADP-ribose) glycohydrolase. Nat. Comm., 3, 878. https://www.doi.org/10.1038/ncomms1889

11- Nehmé, H., Nehmé, R., Lafite, P., Routier, S., & Morin, P. (2012). New development in in-capillary electrophoresis techniques for kinetic and inhibition study of enzymes. Anal. Chim. Acta, 722, 127–135. https://www.doi.org/10.1016/j.aca.2012.02.003

10- Slade, D., Dunstan, M. S., Barkauskaite, E., Weston, R., Lafite, P., Dixon, N., Hergenrother, P. J., Leys, D., Ahel, I. (2011). The structure and catalytic mechanism of a poly(ADP-ribose) glycohydrolase. Nature, 477(7366), 616–620. https://www.doi.org/10.1038/nature10404

9- Driscoll, M. D., McLean, K. J., Levy, C., Mast, N., Pikuleva, I. A., Lafite, P., Rigby, S. E. J., Leys, D., Munro, A. W. (2010). Structural and biochemical characterization of Mycobacterium tuberculosis CYP142. J. Biol. Chem., 285(49), 38270–38282. https://www.doi.org/10.1074/jbc.M110.164293

8- Heyes, D.J., Levy, C., Lafite, P., Roberts, I. S., Goldrick, M., Stachulski, A.V., Rossington, S.B., Stanford, D., Rigby, S.E., Scrutton, N.S., Leys, D. (2009). Structure-based mechanism of CMP-2-keto-3-deoxymanno-octulonic acid synthetase: convergent evolution of a sugar-activating enzyme with DNA/RNA polymerases. J. Biol. Chem., 284(51), 35514–35523. https://www.doi.org/10.1074/jbc.M109.056630

7- McLean, K. J., Lafite, P., Levy, C., Cheesman, M. R., Mast, N., Pikuleva, I. A., Leys, D, Munro, A. W. (2009). The structure of Mycobacterium tuberculosis CYP125 molecular basis for cholesterol binding in a P450 needed for host infection. J. Biol. Chem., 284(51), 35524–35533. https://www.doi.org/10.1074/jbc.M109.032706

6- Pudney, C. R., McGrory, T., Lafite, P., Pang, J. Y., Hay, S., Leys, D., Sutcliffe, M. J. , Scrutton, N. S. (2009). Parallel pathways and free-energy landscapes for enzymatic hydride transfer probed by hydrostatic pressure. Chembiochem, 10(8), 1379–1384. https://www.doi.org/10.1002/cbic.200900071

5- Tralau, T., Lafite, P., Levy, C., Combe, J. P., Scrutton, N. S., Leys, D. (2009). An internal reaction chamber in dimethylglycine oxidase provides efficient protection from exposure to toxic formaldehyde. J. Biol. Chem., 284(26), 17826–17834. https://www.doi.org/ 10.1074/jbc.M109.006262

4- Lafite, P., André, F., Zeldin, D. C., Dansette, P. M., Mansuy, D. (2007). Unusual regioselectivity and active site topology of human cytochrome P450 2J2. Biochemistry, 46(36), 10238–10247. https://www.doi.org/10.1021/bi700876a

3- Lafite, P., Dijols, S., Zeldin, D. C., Dansette, P. M., Mansuy, D. (2007). Selective, competitive and mechanism-based inhibitors of human cytochrome P450 2J2. Arch. Biochem. Biophys., 464(2), 155–168. https://www.doi.org/10.1016/j.abb.2007.03.028

2- Lafite, P., Dijols, S., Buisson, D., Macherey, A. C., Zeldin, D. C., Dansette, P. M., Mansuy, D. (2006). Design and synthesis of selective, high-affinity inhibitors of human cytochrome P450 2J2. Bioorg. Med. Chem. Lett., 16(10), 2777–2780. https://www.doi.org/10.1016/j.bmcl.2006.02.004

1- Pfister, S. L., Spitzbarth, N., Zeldin, D. C., Lafite, P., Mansuy, D., Campbell, W. B. (2003). Rabbit aorta converts 15-HPETE to trihydroxyeicosatrienoic acids: potential role of cytochrome P450. Arch. Biochem. Biophys., 420(1), 142–152. https://doi.org/10.1016/j.abb.2003.09.026

Patent:

1- Yaacoub K., Guillaudeux T., Daniellou R., Lafite P., Aci-Sèche S., Bonnet P., Selective c-FLIP Inhibitors as anticancer agents. Eur. Pat. EP 3323428 (23/05/18); Int. Pat. WO 2018/091647 (24/05/18).

Reviews:

3- Ati, J, Lafite P., Daniellou R (2017). Enzymatic synthesis of glycosides: from natural O- and N-glycosides to rare C- and S-glycosides. Belstein J. Org. Chem., 13, 1857-1865. https://www.doi.org/10.3762/bjoc.13.180

2- Lafite, P., Daniellou, R. (2012). Rare and unusual glycosylation of peptides and proteins. Nat. Prod. Rep., 29(7), 729–738. https://www.doi.org/10.1039/c2np20030a

1- Mansuy, D., Lafite, P. (2007). Great adaptability of the heme-cysteinate monooxygenases family to very diverse substrates and sophisticated reactions. J Porphyr. Phtalocyan., 11(3–4), 258–268. https://www.doi.org/ 10.1142/S108842460700031X

Book Chapters:

2- Peyrot C., Lafite, P., Lemiègre L., Daniellou, R. (2017) Low molecular weight carbohydrate-based hydrogelators In Carbohydrate Chemistry: Volume 43 (Vol. 43, pp. 245-265). The Royal Society of Chemistry. https://www.doi.org/ 10.1039/9781788010641-00245

1- Guillotin, L., Lafite, P., Daniellou, R. (2014). Chapter 10 : Enzymatic thioglycosylation: current knowledge and challenges. In Carbohydrate Chemistry: Volume 40 (Vol. 40, pp. 178–194). The Royal Society of Chemistry. https://www.doi.org/ 10.1039/9781849739986-00178