title: Accurate Determination of Conformational Transitions in Oligomeric Membrane Proteins.
abstract: The structural dynamics governing collective motions in oligomeric membrane proteins play key roles in vital biomolecular processes at cellular membranes. In this study, we present a structural refinement approach that combines solid-state NMR experiments and molecular simulations to accurately describe concerted conformational transitions identifying the overall structural, dynamical, and topological states of oligomeric membrane proteins. The accuracy of the structural ensembles generated with this method is shown to reach the statistical error limit, and is further demonstrated by correctly reproducing orthogonal NMR data. We demonstrate the accuracy of this approach by characterising the pentameric state of phospholamban, a key player in the regulation of calcium uptake in the sarcoplasmic reticulum, and by probing its dynamical activation upon phosphorylation. Our results underline the importance of using an ensemble approach to characterise the conformational transitions that are often responsible for the biological function of oligomeric membrane protein states.
authors.affiliation: Department of Life Sciences, Imperial College London, South Kensington, London, SW7 2AZ, UK.
authors.forename: Máximo
authors.initials: M
authors.lastname: Sanz-Hernández
authors.affiliation: Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Vitaly V
authors.initials: VV
authors.lastname: Vostrikov
authors.affiliation: Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, MN 55455, USA.|Department of Chemistry, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Gianluigi
authors.initials: G
authors.lastname: Veglia
authors.affiliation: Department of Life Sciences, Imperial College London, South Kensington, London, SW7 2AZ, UK.
authors.forename: Alfonso
authors.initials: A
authors.lastname: De Simone
mesh_terms: D000465:Algorithms; D000818:Animals; D016880:Anisotropy; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism; D003198:Computer Simulation; D006801:Humans; D008051:Lipid Bilayers / Q000737:chemistry* / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism; D056004:Molecular Dynamics Simulation; D010766:Phosphorylation; D011487:Protein Conformation; D055503:Protein Multimerization; D000099211:Phospholamban
chemical_list: D002135:Calcium-Binding Proteins; D008051:Lipid Bilayers; D008565:Membrane Proteins; D000099211:Phospholamban