Solid-state deuterium NMR spectroscopy of rhodopsin

Suchithranga M.D.C. Perera, Xiaolin Xu, Trivikram R. Molugu, Andrey V. Struts, Michael F Brown

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Rhodopsin is a prototype for the large Family A of G-protein-coupled receptors (GPCRs). These proteins regulate many signaling processes, and more than 35% of human pharmaceuticals are targeted against diseases related to dysfunctions of GPCR pathways. Membrane proteins such as GPCRs are challenging to crystallize for X-ray studies. In addition, their effective molar masses in detergent solutions push the limits for solution NMR spectroscopy. By contrast, solidstate NMR allows both the structure and dynamics of membrane proteins to be investigated in a natural lipid bilayer environment. Here, we describe solid-state 2H NMR methods for investigating structural and dynamical changes of the retinylidene cofactor of the GPCR rhodopsin upon photoillumination. Rhodopsin was regenerated with retinal containing 2H-labeled C5-, C9-, or C13-methyl groups. The receptor was recombined with phospholipid membranes, which were aligned on planar glass slides. The angular dependences of the 2H NMR spectra and the corresponding relaxation rates were measured for rhodopsin in the dark and in the cryo-trapped, preactive Meta-I and active Meta-II states. Analysis of the 2H NMR lineshapes using a static uniaxial distribution yields orientational restraints for the retinylidene conformation when bound to the protein. Solid-state 2H NMR relaxation data provide additional information on the motion of the bound cofactor. The structural and dynamical changes of retinal reveal howits functional groups (methyl groups and the ß-ionone ring) affect rhodopsin light activation, and illustrate the opportunities of solid-state 2H NMR spectroscopy in studying membrane proteins.

Original languageEnglish (US)
Title of host publicationModern Magnetic Resonance
PublisherSpringer International Publishing
Pages1251-1270
Number of pages20
ISBN (Electronic)9783319283883
ISBN (Print)9783319283876
DOIs
StatePublished - Jun 13 2018

Keywords

  • GPCRs
  • Lipids
  • Membrane proteins
  • Membranes
  • Molecular dynamics
  • NMR relaxation
  • NMR spectroscopy
  • Rhodopsin activation

ASJC Scopus subject areas

  • Engineering(all)
  • Materials Science(all)
  • Medicine(all)
  • Social Sciences(all)
  • Arts and Humanities(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

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  • Cite this

    Perera, S. M. D. C., Xu, X., Molugu, T. R., Struts, A. V., & Brown, M. F. (2018). Solid-state deuterium NMR spectroscopy of rhodopsin. In Modern Magnetic Resonance (pp. 1251-1270). Springer International Publishing. https://doi.org/10.1007/978-3-319-28388-3_144