title: Quantifying the stabilizing effects of protein-ligand interactions in the gas phase.
abstract: The effects of protein-ligand interactions on protein stability are typically monitored by a number of established solution-phase assays. Few translate readily to membrane proteins. We have developed an ion-mobility mass spectrometry approach, which discerns ligand binding to both soluble and membrane proteins directly via both changes in mass and ion mobility, and assesses the effects of these interactions on protein stability through measuring resistance to unfolding. Protein unfolding is induced through collisional activation, which causes changes in protein structure and consequently gas-phase mobility. This enables detailed characterization of the ligand-binding effects on the protein with unprecedented sensitivity. Here we describe the method and software required to extract from ion mobility data the parameters that enable a quantitative analysis of individual binding events. This methodology holds great promise for investigating biologically significant interactions between membrane proteins and both drugs and lipids that are recalcitrant to characterization by other means.
authors.affiliation: Department of Chemistry, University of Oxford, South Parks Road, Oxford OX1 5QY, UK.
authors.forename: Timothy M
authors.initials: TM
authors.lastname: Allison
authors.affiliation: Department of Chemistry, University of Oxford, South Parks Road, Oxford OX1 5QY, UK.
authors.forename: Eamonn
authors.identifier: http://orcid.org/0000-0001-8219-0052
authors.initials: E
authors.lastname: Reading
authors.affiliation: Department of Chemistry, University of Oxford, South Parks Road, Oxford OX1 5QY, UK.
authors.forename: Idlir
authors.initials: I
authors.lastname: Liko
authors.affiliation: Department of Chemistry, University of Oxford, South Parks Road, Oxford OX1 5QY, UK.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Baldwin
authors.affiliation: Department of Chemistry, University of Oxford, South Parks Road, Oxford OX1 5QY, UK.
authors.forename: Arthur
authors.initials: A
authors.lastname: Laganowsky
authors.affiliation: Department of Chemistry, University of Oxford, South Parks Road, Oxford OX1 5QY, UK.
authors.forename: Carol V
authors.initials: CV
authors.lastname: Robinson
mesh_terms: D006801:Humans; D008024:Ligands; D013058:Mass Spectrometry; D008958:Models, Molecular; D011485:Protein Binding; D055550:Protein Stability; D058767:Protein Unfolding; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D008024:Ligands; D011506:Proteins