Investigation of rhodopsin dynamics in its signaling state by solid-state deuterium NMR spectroscopy

Andrey V. Struts, Udeep Chawla, Suchithranga M D C Perera, Michael F Brown

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Scopus citations

Abstract

Site-directed deuterium NMR spectroscopy is a valuable tool to study the structural dynamics of biomolecules in cases where solution NMR is inapplicable. Solid-state 2H NMR spectral studies of aligned membrane samples of rhodopsin with selectively labeled retinal provide information on structural changes of the chromophore in different protein states. Moreover 2H NMR relaxation time measurements allow one to study the dynamics of the ligand during the transition from the inactive to the active state. Here we describe the methodological aspects of solid-state 2H NMR spectroscopy for functional studies of rhodopsin, with an emphasis on the dynamics of the retinal cofactor. We provide complete protocols for the preparation of NMR samples of rhodopsin with 11-cis-retinal selectively deuterated at the methyl groups in aligned membranes. In addition we review optimized conditions for trapping the rhodopsin photointermediates; and we address the challenging problem of trapping the signaling state of rhodopsin in aligned membrane films.

Original languageEnglish (US)
Title of host publicationRhodopsin: Methods and Protocols
PublisherSpringer New York
Pages133-158
Number of pages26
ISBN (Print)9781493923304, 9781493923298
DOIs
StatePublished - Feb 19 2015

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Keywords

  • G protein-coupled receptor
  • Lipids
  • Membrane
  • Nuclear magnetic resonance
  • Protein dynamics
  • Relaxation
  • Rhodopsin
  • Vision

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Struts, A. V., Chawla, U., Perera, S. M. D. C., & Brown, M. F. (2015). Investigation of rhodopsin dynamics in its signaling state by solid-state deuterium NMR spectroscopy. In Rhodopsin: Methods and Protocols (pp. 133-158). Springer New York. https://doi.org/10.1007/978-1-4939-2330-4_10