Functional Studies on Membrane Proteins by Means of H/D Exchange in Infrared: Structural Changes in Na+ NQR from V. cholerae in the Presence of Lipids

  • Yashvin Neehaul
  • Sebastien Kriegel
  • Blanca Barquera
  • Petra Hellwig
Part of the Methods in Molecular Biology book series (MIMB, volume 1635)


H/D exchange kinetics at the level of the amide proton in the mid infrared (1700–1500 cm−1) make it possible to study the conformational flexibility of membrane proteins, independent of size or the presence of detergent or lipids. Slow, medium, and fast exchanging domains are distinguished, which reveal a different accessibility to the solvent. Whereas amide hydrogens undergo rapid exchange with solvent in an open structure, hydrogens experience much slower exchange when involved in H-bonded structures or when sterically inaccessible to the solvent. Here, we describe the protocol that was used to study the effect of phospholipids on the overall structure of the Na+ NQR from V. cholerae, a sodium pumping membrane protein.

Key words

H/D exchange kinetics Secondary structure analysis Infrared spectroscopy 



We are grateful to the University of Strasbourg, the CNRS, and the foundation FRC for ongoing financial support.


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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Yashvin Neehaul
    • 1
    • 2
  • Sebastien Kriegel
    • 1
  • Blanca Barquera
    • 3
  • Petra Hellwig
    • 1
  1. 1.Laboratoire de Bioelectrochimie et Spectroscopie, UMR 7140, Chimie de la Matière ComplexeUniversité de StrasbourgStrasbourgFrance
  2. 2.Mauritius Oceanography InstituteAlbionMauritius
  3. 3.Department of Biological Sciences, Center for Biotechnology and Interdisciplinary StudiesRensselaer Polytechnic InstituteTroyUSA

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