title: Using Hydrogen-Deuterium Exchange Mass Spectrometry to Examine Protein-Membrane Interactions.
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.
authors.affiliation: Pharmaceutical Sciences Section, University of Geneva, Geneva, Switzerland.
authors.forename: O
authors.initials: O
authors.lastname: Vadas
authors.affiliation: University of Victoria, Victoria BC, Canada.
authors.forename: M L
authors.initials: ML
authors.lastname: Jenkins
authors.affiliation: University of Victoria, Victoria BC, Canada.
authors.forename: G L
authors.initials: GL
authors.lastname: Dornan
authors.affiliation: University of Victoria, Victoria BC, Canada. Electronic address: jeburke@uvic.ca.
authors.forename: J E
authors.initials: JE
authors.lastname: Burke
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D058534:Class I Phosphatidylinositol 3-Kinases; D041961:Deuterium Exchange Measurement / Q000379:methods*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D009154:Mutation; D051059:PTEN Phosphohydrolase / Q000235:genetics / Q000378:metabolism*; D019869:Phosphatidylinositol 3-Kinases / Q000235:genetics / Q000378:metabolism*; D010716:Phosphatidylinositols / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D025941:Protein Interaction Mapping; D011499:Protein Processing, Post-Translational; D015398:Signal Transduction
chemical_list: D058534:Class I Phosphatidylinositol 3-Kinases; D008565:Membrane Proteins; D051059:PTEN Phosphohydrolase; D019869:Phosphatidylinositol 3-Kinases; D010716:Phosphatidylinositols; C421683:Pik3cd protein, mouse; C494929:PTEN protein, human
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