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About

The Hydrogen/Deuterium eXchange Mass Spectrometry activity at Institut Pasteur

 

Hydrogen Deuterium eXchange followed by Mass Spectrometry (HDX-MS) is a powerful and recognized biophysical tool in structural biology capable of probing protein/ligand interactions, conformational changes, and protein folding and dynamics (Figure 1). HDX-MS measures the changes in mass imposed by the replacement of backbone amide hydrogens by deuterium added in excess in the surrounding environment. The rate of exchange relies on the folding and dynamics of proteins making backbone amide hydrogens excellent structural probes (more details about the principle of hydrogen exchange can be found here).

 

 

We  are currently using an optimized version of the HDX-MS pipeline  commercialized by the Waters Company composed of a Synapt G2-Si HDMS mass spectrometer with ETD and IMS capabilities, a LEAP-Pal robot for automated sample handling, a cooled HDX manager that maintains the valves, columns and associated tubings at 0°C and Class M nanoACQUITY UPLC pumps (Fundings: CACSICE Equipex, C2RT Institut Pasteur). Our pipeline has been completed with the development of a software named “MEMHDX” to aid in the rapid statistical validation and visualization of large HDX-MS datasets. A schematic overview of a  classical HDX-MS experiment performed with our optimized HDX-MS  workflow is presented in Figure 2.

 

 

The majority of our projects focus on soluble, membrane-associated or integral membrane proteins that are either involved in infectious disease or considered as potential drug targets. Most of these projects are carried out in collaboration with internal or external collaborators. Our  expertise in HDX-MS include:

  • Analysis of the conformation and dynamic of intrinsically disordered proteins (IDPs) and large proteins (> 150 kDa).
  • Analysis of small molecule/protein interactions.
  • Analysis of protein/protein interaction (e.g., activation of the CyaA toxin) and identification of binding sites (e.g. Epitope mapping for both the academic and pharmaceutical sector).
  • Analysis of protein/DNA interactions.
  • Analysis of the conformation and dynamic of integral membrane proteins (e.g., the human glutamate transporteur EAAT1).
  • Optimization and improvement of the HDX-MS pipeline (e.g. MEMHDX; details about the principle of the software can be found here).

 

Main Projects and Collaborators

  • Structural analysis of B. pertussis CyaA toxin – Collaboration with the group of Alexandre Chenal, Institut Pasteur, Paris.
  • Structural analysis of viral IDPs – Collaboration with the group of Sonia Longhi, CNRS, Marseille.
  • Epitope mapping  – Collaboration with Michel-Robert Popoff and Alexandre Chenal, Institut Pasteur, Paris.
  • Deciphering the structural changes associated with ion binding on the human glutamate transporter – Collaboration with the group of Nicolas Reyes, Institut Pasteur, Paris.
  • Identification of protein/aptamer binding sites by HDX-MS – Collaboration with the groupe of Marcel Hollenstein, Institut Pasteur, Paris.

 

For more information, you can contact us at: sebastien.brier@pasteur.fr

Events

HDX-MS/NMR DAY

Room: Amphitheater François Jacob – Address: Institut Pasteur, Rue du Docteur Roux, Paris, France
2017-03-16 09:00:00 2017-03-16 18:00:00 Europe/Paris HDX-MS/NMR DAY Dear Colleagues, We are pleased to invite you to attend our upcoming Hydrogen Deuterium eXchange Day (HDX-Day) that will be held at the Institut Pasteur, Paris, on Thursday March 16th 2017. The aim of this […] Institut Pasteur, Rue du Docteur Roux, Paris, France

Projects

CV

Sébastien Brier  holds an Engineer degree in Biology from Polytech Clermont-Ferrand, and received his Ph.D. in Structural Proteomics at the group of Dr. Eric Forest in 2005 in Grenoble, developing HDX-MS approaches to study the binding of small inhibitors to human mitotic kinesins. He completed postdoctoral work at the Barnett Institute of Chemical Biology in Boston in the group of Professor John. R. Engen, and at the CNRS in Paris, France.

In 2009, he was appointed research investigator at Novartis Vaccines and Diagnostics in Siena, Italy, to create a HDX-MS platform dedicated to the structural analysis of protein vaccines (Epitope mapping) . In Autumn 2012 he came back to Paris as a research associate in Structural Proteomics at Stallergenes-Greer, working on the use of the HDX technology to map IgE epitopes on natural allergens to optimize the design of recombinant vaccines. Currently, Sébastien has implemented and manages the HDX-MS activity at Institut Pasteur in Paris working on membrane-associated proteins involved in infectious disease.

Publications

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