Accurate Determination of Conformational Transitions in Oligomeric Membrane Proteins

Sci Rep. 2016 Mar 15:6:23063. doi: 10.1038/srep23063.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Animals
  • Anisotropy
  • Calcium-Binding Proteins / chemistry
  • Calcium-Binding Proteins / metabolism
  • Cell Membrane / chemistry*
  • Cell Membrane / metabolism
  • Computer Simulation
  • Humans
  • Lipid Bilayers / chemistry*
  • Lipid Bilayers / metabolism
  • Magnetic Resonance Spectroscopy / methods
  • Membrane Proteins / chemistry*
  • Membrane Proteins / metabolism
  • Molecular Dynamics Simulation
  • Phosphorylation
  • Protein Conformation*
  • Protein Multimerization*

Substances

  • Calcium-Binding Proteins
  • Lipid Bilayers
  • Membrane Proteins
  • phospholamban