Paulette Charlier

Professor

Detailed information

Publication list

Fréderic Kerff

Research Associate FRS_FNRS

Detailed information

Publication list

Research interest

  • Protein structures: x-ray crystallography, modelling, docking
  • Membrane protein crystallography
  • Protein-ligand interaction (small molecules, RNA/DNA, proteins)
  • Bacterial cell wall peptidoglycan metabolism
  • Bacterial resistance to β-lactam antibiotics

Research summary

Our activities are dedicated to the determination and analysis of the 3D structures of soluble as well as membrane proteins. Our analytical method is X-ray crystallography with the use of the last breakthroughs of the domain: production of seleno-methionyl proteins for phasing and data collection at different synchrotron beamlines. Our laboratory benefits from access to the Protein Factory and Robotein platforms of CIP for production, purification and biochemical/biophysical analysis of proteins, a Mosquito robot for automated screening of crystallization conditions, and ESRF (Grenoble, France) and SOLEIL (Saint-Aubin, France) beamlines for X-ray data collection.

In order to understand the function and/or activity of a protein, we combine the structural analysis with information resulting from other biophysical and biochemical methods. Interactions of a protein with its potential ligand (ions, small organic molecules, nucleic acids, proteins) could be investigated by experimental methods (Microscale Thermophoresis, Surface Plasmon Resonance, Fragment Based Screening by X-ray crystallography) and theoretical ones (molecular modeling, docking) as well.

The first main thematic research concerns the metabolism of the peptidoglycan (PG), the essential component of bacterial cell wall. Its metabolism is an essentially dynamic process during which are involved numerous enzymes which could be many potential targets for new approaches in antibiotic search. Amongst the enzymes studied are the transglycosylases and transpeptidases, as monofunctional enzymes or part of multi-domain proteins called PBPs (Penicillin-binding proteins), endopeptidases, amidases and muramidases.

The second main thematic research concerns proteins involved in the bacterial resistance phenomenon, in particular developed against b-lactam antibiotics. Amongst the enzymes studied are the b-lactamases (from classes A, B, C and D) and the PBPs that have emerged in resistant strains.

Our research also includes the characterization of undecaprenyl-pyrophosphate (C55-PP) phosphatases, the membrane proteins involved in the synthesis and recycling of the lipid carrier used for the peptidoglycan polymerization. C55-PP phosphatases belong to two families: the PAP2 and BacA/UppP families. Several enzymes from these families are present in a single bacterium raising the question of the significance of this redundancy. In this context, we are studying the proteins from E. coli, Bacillus subtilis and the human pathogen Helicobacter pylori. In addition to the C55-PP phosphatase activity, some PAP2 enzymes are also involved in the phosphatidyglycerol and lipid A metabolisms.

Lab members

  • Eric Sauvage (Postdoc)
  • Raphael Herman (Technician)
  • François Delbrassine (PhD student)
  • Raphael Léonard (PhD student)

Selected publications

  1. Sauvage Eric, Kerff, Frédéric, Terrak Mohammed, Ayala Juan & Charlier Paulette. (2008) The Penicillin-Binding Proteins: Structure and Role in Peptidoglycan Biosynthesis. FEMS Microbiology Reviews 32, 234-258.
  1. Kerff F., Amoroso A., Herman R., Sauvage E., Petrella S., Filée P., Charlier P., Joris B., Tabuchi A., Nikolaidis N., Cosgrove D.J. (2008) Crystal structure and activity of Bacillus subtilis YoaJ (EXLX1), a bacterial expansin that promotes root colonization. Proc Natl Acad Sci U S A. 105, 16876-81.
  1. Eric Sauvage, Astrid Zervosen, Georges Dive, Raphael Herman, Ana Amoroso, Bernard Joris, Eveline Fonzé, F. Pratt, André Luxen, Paulette Charlier & Frédéric Kerff (2009) Structural basis of the inhibition of class A b-lactamases and penicillin-binding proteins by 6-b-iodopenicillanate. J.Am.Chem.Soc. 131, 15262–15269.
  1. Frédéric Kerff, Stéphanie Petrella, Frédéric Mercier, Eric Sauvage, Raphaël Herman, Anne Pennartz, Astrid Zervosen, André Luxen, Jean-Marie Frère, Bernard Joris & Paulette Charlier (2010) Specific Structural Features of the N-Acetylmuramoyl-L-Alanine Amidase AmiD from Escherichia coli and Mechanistic Implications for Enzymes of This Family. Mol. Biol. 397, 249–259
  1. Rocaboy M, Herman R, Sauvage E, Remaut H, Moonens K, Terrak M, Charlier P, Kerff F (2013) The crystal structure of the cell division amidase AmiC reveals the fold of the AMIN domain, a new peptidoglycan binding domain. Microbiol. 90, 267-277.
  1. Eric Sauvage, Adeline Derouaux, Claudine Fraipont, Marine Joris, Raphaël Herman, Mathieu Rocaboy, Marie Schloesser, Jacques Dumas, Frédéric Kerff, Martine Nguyen-Distèche & Paulette Charlier (2014) Crystal structure of penicillin-binding protein 3 (PBP3) from Escherichia coli. Plos One. 9(5)