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Probing Protein–Membrane Interactions and Dynamics Using Hydrogen–Deuterium Exchange Mass Spectrometry (HDX-MS)

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Protein-Ligand Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2263))

Abstract

Cellular membranes are a central hub for initiation and execution of many signaling processes. Integral to these processes being accomplished appropriately is the highly controlled recruitment and assembly of proteins at membrane surfaces. The study of the molecular mechanisms that mediate protein–membrane interactions can be facilitated by utilizing hydrogen–deuterium exchange mass spectrometry (HDX-MS). HDX-MS is a robust analytical technique that allows for the measurement of the exchange rate of backbone amide hydrogens with solvent to make inferences about protein structure and conformation. This chapter discusses the use of HDX-MS as a tool to study the conformational changes that occur within peripheral membrane proteins upon association with membrane. Particular reference will be made to the analysis of the protein kinase Akt and its activation upon binding phosphatidylinositol (3,4,5) tris-phosphate (PIP3)-containing membranes to illustrate specific methodological principles.

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Acknowledgments

This work was supported by a Canadian Institutes of Health Research New Investigator award and Open Operating Grant CRN-142393, Cancer Research Society Operating Grants CRS-22641 and CRS-24368, Natural Sciences and Engineering Research Council of Canada Discovery Grants NSERC-2014-05218 and NSERC-2020-04241, and Michael Smith Foundation for Health Research Scholar Award 17686. The authors declare no conflicts of interest with the contents of this chapter.

Author information

Authors and Affiliations

  1. Department of Biochemistry and Microbiology, University of Victoria, Victoria, BC, Canada

    Jordan T. B. Stariha, Reece M. Hoffmann, David J. Hamelin & John E. Burke

  2. Department of Biochemistry and Molecular Biology, The University of British Columbia, Vancouver, BC, Canada

    John E. Burke

Authors
  1. Jordan T. B. Stariha
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  2. Reece M. Hoffmann
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  3. David J. Hamelin
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  4. John E. Burke
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Corresponding author

Correspondence to John E. Burke .

Editor information

Editors and Affiliations

  1. The Institute of Cancer Research, London, UK

    Tina Daviter

  2. MRC Laboratory of Molecular Biology, Cambridge, UK

    Christopher M. Johnson

  3. MRC Laboratory of Molecular Biology, Cambridge, UK

    Stephen H. McLaughlin

  4. ISMB BiophysX Centre, Birkbeck, University of London, London, UK

    Mark A. Williams

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Stariha, J.T.B., Hoffmann, R.M., Hamelin, D.J., Burke, J.E. (2021). Probing Protein–Membrane Interactions and Dynamics Using Hydrogen–Deuterium Exchange Mass Spectrometry (HDX-MS). In: Daviter, T., Johnson, C.M., McLaughlin, S.H., Williams, M.A. (eds) Protein-Ligand Interactions. Methods in Molecular Biology, vol 2263. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1197-5_22

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  • DOI: https://doi.org/10.1007/978-1-0716-1197-5_22

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1196-8

  • Online ISBN: 978-1-0716-1197-5

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