Journal of Biomolecular NMR

, Volume 16, Issue 2, pp 121–125 | Cite as

Measurement of dipolar couplings in a transducin peptide fragment weakly bound to oriented photo-activated rhodopsin

  • Bernd W. Koenig
  • Drake C. Mitchell
  • Simone König
  • Stephan Grzesiek
  • Burton J. Litman
  • Ad Bax


Rhodopsin-containing disks, isolated from rod outer segments of bovine retina, align at high magnetic fields with their membrane normal parallel to the magnetic field. After light-activation of rhodopsin, transient binding of the C-terminal transducin undecapeptide, selectively labeled with 15N at Leu5 and Gly9, results in residual dipolar contributions to the 1JNH splittings for these two residues. Both residues show 1JNH splittings which are smaller than in the dark-adapted or rhodopsin-free sample, and return to their isotropic values at a rate determined by the decay of the meta II state of rhodopsin. The dipolar couplings indicate that in the bound state, N-H vectors of Leu5 and Gly9 make angles of 48±4° and 40±8°, respectively, with the disk normal.  These `transferred' dipolar couplings potentially offer a useful method for studying the conformation and orientation of flexible, low affinity ligands when bound to oriented integral membrane receptors.

alignment membrane protein receptor rhodopsin transferred dipolar coupling transducin 


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Copyright information

© Kluwer Academic Publishers 2000

Authors and Affiliations

  • Bernd W. Koenig
    • 1
  • Drake C. Mitchell
    • 2
  • Simone König
    • 3
  • Stephan Grzesiek
    • 4
  • Burton J. Litman
    • 2
  • Ad Bax
    • 1
  1. 1.Laboratory of Chemical Physics, NIDDKNational Institutes of HealthBethesdaU.S.A.
  2. 2.Laboratory of Membrane Biophysics and Biochemistry, NIAAANational Institutes of HealthRockville
  3. 3.Laboratory of Biophysical Chemistry, NHLBINational Institutes of HealthBethesdaU.S.A.
  4. 4.Structural Biology InstituteResearch Center JülichJülichGermany

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