Download Development of innovative mass spectrometry approaches for the

ED BIO SORBONNE PARIS CITE
Proposition de sujet de thèse à l’appui d’une demande de contrat doctoral 2017-2018
Renseignements relatifs à l’Unité de Recherche :
Label et intitulé : USR 2000 Spectrométrie de Masse pour la Biologie
Nom et prénom du Directeur : Chamot-Rooke Julia
Téléphone :
+ 33 1 40 61 38 59
Télécopie :
Courriel: [email protected]
Renseignements relatifs à l’Equipe :
Nom de l’Equipe d’Accueil : USR 2000 Spectrométrie de Masse pour la Biologie
Nom et prénom du responsable : Chamot-Rooke Julia
Qualité du responsable : Directeur de Recherche CNRS
Téléphone :
+ 33 1 40 61 38 59
Télécopie :
Courriel : [email protected]
Renseignements relatifs au sujet de thèse :
Nom et prénom du Directeur de thèse (HDR) : Chamot-Rooke Julia
Qualité : Directeur de Recherche CNRS
Téléphone :
+ 33 1 40 61 38 59
Télécopie :
Courriel : [email protected]
Titre du sujet proposé :
(En français) Détermination de la structure de complexes protéiques impliqués dans la virulence
bactérienne par de nouvelles approches de spectrométrie de masse à haute résolution
(En anglais) Development of innovative mass spectrometry approaches for the structural analysis of
protein complexes involved in bacterial virulence.
Département (cocher le département correspondant au sujet de thèse qui n’est pas obligatoirement le
vôtre) :




Biologie Cellulaire et moléculaire, Physiologie et Physiopathologie
Immunologie
Développement Génétique Neurobiologie et Vieillissement
Infectiologie, Microbiologie
Summary (5 lines maximum) :
This PhD project proposes the development of new Mass Spectrometry (MS) based approaches for the
structural analysis of challenging protein complexes. Two main axes will be developed: covalent crosslinking combined to MS and native MS. These developments will be used to decipher the structure of
bacterial secretion systems, which are sophisticated nanomachineries involved in bacterial virulence.
Proposition de sujet de thèse à l’appui d’une demande de contrat doctoral 2017-2087
Nom, prénom du directeur de l'unité de recherche : Chamot-Rooke Julia
Numéro de l'unité de recherche (et établissement de rattachement) : USR 2000
Nom, prénom du responsable de l'équipe d'accueil (EAD) : Chamot-Rooke Julia
Nom, prénom du directeur de thèse : Chamot-Rooke Julia
Titre du sujet de thèse proposé (en anglais): Development of innovative mass spectrometry approaches
for the structural analysis of protein complexes involved in bacterial virulence.
5 key words: mass spectrometry, cross-linking, structural biology, protein complexes, bacterial virulence
Candidat pressenti :

OUI

NON
Contenu scientifique du programme de la thèse (en anglais)
Physical interactions between proteins form the basis of most biological functions. Therefore, a critical
step toward understanding cellular processes is to determine the structure of protein complexes and
reveal how their structural organization can relate to their function. However, studying the threedimensional organization of large protein assemblies represents a major challenge for structural
biologists. In the last decade, Mass Spectrometry (MS) has become a very valuable tool for analyzing
such complexes offering an unrivaled sensitivity and the possibility to analyze very large complexes in
their biological state. For instance, cross-linking combined to MS leads to the identification of aminoacids that are in close proximity and thus provide spatial constraints for the modeling1. Our group
recently designed a new trifunctional cross-linker (named NNP9) dedicated to MS analysis, which
outperforms all commercial ones and allows the analysis of large membrane systems2. MS can also be
used to analyze protein complexes in their native state (native MS). For these experiments, the noncovalent interactions between subunits are kept intact and thus the stoichiometry of the complex can be
determined by the accurate measurement of its molecular mass. Additional fragmentation steps can also
be performed to obtain information on the complex topology.
Our goal, with this PhD project is to combine both cross-linking MS and native MS to decipher the
structure of bacterial secretion systems that are challenging protein complexes involved in bacterial
virulence. In a first part, the PhD student will evaluate the efficiency of several newly synthetized crosslinkers. These developments will be performed on model protein complexes. Native mass spectrometry
experiments will be optimized in parallel. In a second part, he/she will work on elucidating the structure of
several bacterial secretion systems in collaboration with different teams of biologists. These protein
complexes are sophisticated nanomachineries anchored in the bacterial membrane and used by many
bacterial pathogens to deliver toxins or other effectors into target cells. The newly developed MS-based
approaches will be used to obtain high quality structural information. All data will finally be combined to
build structural models for the complexes of interest.
The PhD student will have full access to state-of-the-art high-resolution LC-MS mass spectrometers
present in the lab (Orbitrap Lumos, Orbitrap Q-Exactive). He will also participate to high-level
instrumental developments.
References:
(1) Petrotchenko, E. V.; Borchers, C. H. Mass Spectrom. Rev. 2010, 29, 862−876.
(2) Nury, C. et al. 2015, 87, 1853−1860
Indiquez les cinq meilleures publications récentes de l’équipe :
Canul-Tec, J.C. et al. Nature (in press).
Vorontsov, E.A., et al. Mol Cell Proteomics 15, 3388-3404 (2016).
Nury, C., et al. Anal Chem 87, 1853-1860 (2015).
Gault, J., et al. Plos Pathog 11 (2015).
Smith, L.M., et al. Nat Methods 10, 186-187 (2013).