Faculté de Pharmacie
Membrane transport, Antibioresistance, and Signaling
Structural study of membrane proteins
Our team, specialized in the structural study of membrane proteins, focuses its research on the public health problem of antibiotic resistance responsible for the dramatic increase of multi-resistant bacteria all over the word. Among the different ways used by bacteria to resist to antibiotics, the active efflux through tripartite efflux pumps is one of the most important. Our main target is Pseudomonas aeruginosa (PA), a nosocomial opportunistic pathogen that contributes to the decline of respiratory function in cystic fibrosis (CF) patients. PA possesses mainly four efflux pumps proven to be involved in the efflux of antibiotics, with some specificities concerning the family of antibiotics taken in charge. Different strategies can be considered to develop molecules blocking the function of these pumps such as the perturbation of the energy system, the maintaining of the porin in its closed conformation, or the prevention of the assembly. We are conducting this research by complementary approaches linking structural biology (X-ray crystallography, SAXS, Cryo-EM) with in cellulo functional analysis. We recently managed to describe the assembly of the constitutive efflux pump MexA-MexB-OprM by cryo-electronic microscopy in close collaboration with the team of Dr Lambert in Bordeaux University (Glavier, Nat Comm, 2020), making possible a drug design approach.
In parallel, we have initiated different projects:
– the study of the PA efflux pump expression regulators, in close collaboration with the team of Dr C. Llanes (University of Besançon)
– we model the possible effect of single-mutations identified in clinical PA strains isolated from the spits of cystic fibrosis patients in collaboration with the team of Pr P. Plesiat (CHU Besançon, “Centre National de Référence de la Résistance aux Antibiotiques”)
– we evaluate the role of post-translational modifications (PTM) on specific proteins involved in regulation mechanisms, with a particular focus on lysine acetylation, in collaboration with the team of Dr J. Hardouin (University of Rouen).
Structural biology, membrane protein production and purification
Our team is specialized in structural biology, membrane protein production and purification, protein/protein or protein/ligand interaction characterization by different biophysical approaches
We are equipped with a mosquito robot for protein crystallization, a fluorimeter and an ITC
We have a regular access to the synchrotrons localized in France, and to the Pasteur cryo-EM microscope
Top 5 publications
1- Molecular Determinants for OMF Selectivity in Tripartite RND Multidrug Efflux Systems. Boyer E, Dessolin J, Lustig M, Decossas M, Phan G, Cece Q, Durand G, Dubois V, Sansen J, Taveau JC, Broutin I, Daury L, Lambert O. Antibiotics (Basel). 2022;11(2):126. doi: 10.3390/antibiotics11020126.
2- MexAB-OprM Efflux Pump Interaction with the Peptidoglycan of Escherichia coli and Pseudomonas aeruginosa. Ma M, Lustig M, Salem M, Mengin-Lecreulx D, Phan G, Broutin I. Int J Mol Sci. 2021;22(10):5328. doi: 10.3390/ijms22105328.
3- Antibiotic export by MexB multidrug efflux transporter is allosterically controlled by a MexA-OprM chaperone-like complex. Glavier M, Puvanendran D, Salvador D, Decossas M, Phan G, Garnier C, Frezza E, Cece Q, Schoehn G, Picard M, Taveau JC, Daury L, Broutin I*, Lambert O*. Nat Commun. 2020;11(1):4948. doi: 10.1038/s41467-020-18770-5.
4- Functional Mechanism of the Efflux Pumps Transcription Regulators From Pseudomonas aeruginosa Based on 3D Structures. Housseini B Issa K, Phan G, Broutin I. Front Mol Biosci. 2018;5:57. doi: 10.3389/fmolb.2018.00057. eCollection 2018.
5- Xenon for tunnelling analysis of the efflux pump component OprN. Ntsogo Enguéné YV, Phan G, Garnier C, Ducruix A, Prangé T, Broutin I. PLoS One. 2017;12(9):e0184045. doi: 10.1371/journal.pone.0184045. eCollection 2017.
Dernières publications sur HAL :
- [hal-04312576] Purification des protéines membranaires en microfluidiqueby email@example.com (Lucas Lecorgne) on 30 November 2023 at 12h44
Le système nerveux central (SNC) est une cible thérapeutique pour de nombreux composés pharmacologiquement actifs. Pour atteindre leur cible, ces […]
- [hal-04023779] LC–MS/MS-based quantification of efflux transporter proteins at the BBBby firstname.lastname@example.org (David Gomez-Zepeda) on 10 March 2023 at 15h39
- [hal-03815598] The Structural and Functional Study of Efflux Pumps Belonging to the RND Transporters Family from Gram-Negative...by email@example.com (Attilio Vittorio Vargiu) on 14 October 2022 at 17h12
- [hal-03815594] Molecular Determinants for OMF Selectivity in Tripartite RND Multidrug Efflux Systemsby firstname.lastname@example.org (Esther Boyer) on 14 October 2022 at 17h10
Tripartite multidrug RND efflux systems made of an inner membrane transporter, an outer membrane factor (OMF) and a periplasmic adaptor protein (PAP) […]