title: Probing the dynamic regulation of peripheral membrane proteins using hydrogen deuterium exchange-MS (HDX-MS).
abstract: Many cellular signalling events are controlled by the selective recruitment of protein complexes to membranes. Determining the molecular basis for how lipid signalling complexes are recruited, assembled and regulated on specific membrane compartments has remained challenging due to the difficulty of working in conditions mimicking native biological membrane environments. Enzyme recruitment to membranes is controlled by a variety of regulatory mechanisms, including binding to specific lipid species, protein-protein interactions, membrane curvature, as well as post-translational modifications. A powerful tool to study the regulation of membrane signalling enzymes and complexes is hydrogen deuterium exchange-MS (HDX-MS), a technique that allows for the interrogation of protein dynamics upon membrane binding and recruitment. This review will highlight the theory and development of HDX-MS and its application to examine the molecular basis of lipid signalling enzymes, specifically the regulation and activation of phosphoinositide 3-kinases (PI3Ks).
authors.affiliation: Department of Pharmaceutical Chemistry, University of Geneva, 1211 Geneva 4, Switzerland.
authors.forename: Oscar
authors.initials: O
authors.lastname: Vadas
authors.affiliation: Department of Biochemistry and Microbiology, University of Victoria, Victoria, Canada, BC V8P 5C2 jeburke@uvic.ca.
authors.forename: John E
authors.initials: JE
authors.lastname: Burke
mesh_terms: D000818:Animals; D000846:Anniversaries and Special Events; D001363:Awards and Prizes; D001671:Biochemistry; D002462:Cell Membrane / Q000737:chemistry / Q000201:enzymology / Q000378:metabolism*; D058534:Class I Phosphatidylinositol 3-Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D041961:Deuterium Exchange Measurement; D006801:Humans; D013058:Mass Spectrometry; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D009154:Mutation; D011487:Protein Conformation; D017510:Protein Folding; D021381:Protein Transport; D015398:Signal Transduction; D012958:Societies, Scientific; D006113:United Kingdom
chemical_list: D008565:Membrane Proteins; D058534:Class I Phosphatidylinositol 3-Kinases
keywords: catalytic subunit of PI3Kα (PIK3CA); hydrogen deuterium exchange–mass spectrometry (HDX–MS); membrane proteins; p85α regulatory subunit of class I PI3K (PIK3R1); phosphoinositide 3-kinases (PI3Ks); protein dynamics