Using Hydrogen-Deuterium Exchange Mass Spectrometry to Examine Protein-Membrane Interactions

Methods Enzymol. 2017:583:143-172. doi: 10.1016/bs.mie.2016.09.008. Epub 2016 Oct 18.

Abstract

Many fundamental cellular processes are controlled via assembly of a network of proteins at membrane surfaces. The proper recruitment of proteins to membranes can be controlled by a wide variety of mechanisms, including protein lipidation, protein-protein interactions, posttranslational modifications, and binding to specific lipid species present in membranes. There are, however, only a limited number of analytical techniques that can study the assembly of protein-membrane complexes at the molecular level. A relatively new addition to the set of techniques available to study these protein-membrane systems is the use of hydrogen-deuterium exchange mass spectrometry (HDX-MS). HDX-MS experiments measure protein conformational dynamics in their native state, based on the rate of exchange of amide hydrogens with solvent. This review discusses the use of HDX-MS as a tool to identify the interfaces of proteins with membranes and membrane-associated proteins, as well as define conformational changes elicited by membrane recruitment. Specific examples will focus on the use of HDX-MS to examine how large macromolecular protein complexes are recruited and activated on membranes, and how both posttranslational modifications and cancer-linked oncogenic mutations affect these processes.

Keywords: Allosteric modifications; Biophysics; Conformational changes; Fluorescence-resonance energy transfer; GPCR; Hydrogen–deuterium exchange mass spectrometry; Lipid signaling; Membrane proteins; PI3K; Phosphoinositides; Phospholipids; Protein dynamics; Protein–membrane interactions.

Publication types

  • Review

MeSH terms

  • Animals
  • Cell Membrane / chemistry
  • Cell Membrane / metabolism
  • Class I Phosphatidylinositol 3-Kinases
  • Deuterium Exchange Measurement / methods*
  • Humans
  • Mass Spectrometry / methods
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Mice
  • Mutation
  • PTEN Phosphohydrolase / genetics
  • PTEN Phosphohydrolase / metabolism*
  • Phosphatidylinositol 3-Kinases / genetics
  • Phosphatidylinositol 3-Kinases / metabolism*
  • Phosphatidylinositols / metabolism
  • Protein Binding
  • Protein Conformation
  • Protein Interaction Mapping
  • Protein Processing, Post-Translational*
  • Signal Transduction

Substances

  • Membrane Proteins
  • Phosphatidylinositols
  • Class I Phosphatidylinositol 3-Kinases
  • Pik3cd protein, mouse
  • PTEN Phosphohydrolase
  • PTEN protein, human

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