Hydrogen/Deuterium Exchange Mass Spectrometry for the Structural Analysis of Detergent-Solubilized Membrane Proteins

  • Darragh P. O’Brien
  • Véronique Hourdel
  • Alexandre Chenal
  • Sébastien BrierEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2127)


Integral membrane proteins are involved in numerous biological functions and represent important drug targets. Despite their abundance in the human proteome, the number of integral membrane protein structures is largely underrepresented in the Protein Data Bank. The challenges associated with the biophysical characterization of such biological systems are well known. Most structural approaches, including X-ray crystallography, SAXS, or mass spectrometry (MS), require the complete solubilization of membrane proteins in aqueous solutions. Detergents are frequently used for this task, but may interfere with the analysis, as is the case with MS. The use of “MS-friendly” detergents, such as non-ionic alkyl glycoside detergents, has greatly facilitated the analysis of detergent-solubilized membrane proteins. Here, we describe a protocol, which we have successfully implemented in our laboratory to study the structure and dynamics of detergent-solubilized integral membrane proteins by Hydrogen/Deuterium eXchange and Mass Spectrometry (HDX-MS). The procedure does not require detergent removal prior to MS analysis, instead taking advantage of the ultra-high pressure chromatographic system to separate deuterated peptides from “MS-friendly” detergents.

Key words

Integral membrane proteins “MS-friendly” detergents Ligand binding Deuterium exchange Mass spectrometry 



The HDX-MS system was financed by the Equipex CACSICE (ANR-11-EQPX-0008). SB would like to thank Alexandre Chenal, Nadia Izadi-Prunyere, Mariette Matondo, Florence Cordier, Thierry Lang, Daniel Ladant, Iñaki Guijarro and Michael Nilges for their kindness and constant support over the last year.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Darragh P. O’Brien
    • 1
    • 2
  • Véronique Hourdel
    • 3
  • Alexandre Chenal
    • 1
  • Sébastien Brier
    • 4
    Email author
  1. 1.Biochemistry of Macromolecular Interaction Unit, Department of Structural Biology and ChemistryInstitut Pasteur, CNRS UMR3528ParisFrance
  2. 2.Nuffield Department of Medicine, Target Discovery InstituteUniversity of OxfordOxfordUK
  3. 3.Environment and Infectious Risks Unit, Department of Infection and EpidemiologyInstitut PasteurParisFrance
  4. 4.Biological NMR Technological Platform, Center for Technological Resources and Research, Department of Structural Biology and ChemistryInstitut Pasteur, CNRS UMR3528ParisFrance

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