Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Observations of light-induced structural changes of retinal within rhodopsin

Nature volume 405, pages 810–813 (2000)Cite this article

Abstract

Photo-isomerization of the 11-cis retinal chromophore activates the mammalian light-receptor rhodopsin1, a representative member of a major superfamily of transmembrane G-protein-coupled receptor proteins (GPCRs) responsible for many cell signal communication pathways. Although low-resolution (5 Å) electron microscopy studies2,3 confirm a seven transmembrane helix bundle as a principal structural component of rhodopsin, the structure of the retinal within this helical bundle is not known in detail. Such information is essential for any theoretical or functional understanding of one of the fastest occurring photoactivation processes in nature, as well as the general mechanism behind GPCR activation4,5,6. Here we determine the three-dimensional structure of 11-cis retinal bound to bovine rhodopsin in the ground state at atomic level using a new high-resolution solid-state NMR method7. Significant structural changes are observed in the retinal following activation by light to the photo-activated MI state of rhodopsin giving the all-trans isomer of the chromophore. These changes are linked directly to the activation of the receptor, providing an insight into the activation mechanism of this class of receptors at a molecular level.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: In the magic angle oriented sample spinning (MAOSS) NMR approach used here, uni-axially oriented phospholipid membranes containing bovine rhodopsin were stacked on glass plates in a MAS rotor, with rotor axis ZR parallel to the sample director.
Figure 2: Deuterium-MAOSS NMR spectra of [18-C(2H)3]-retinal in dark adapted rhodopsin (a) and upon photo-activation in the M I state (illuminated below 273 K) (b), at ω r = 2860 Hz and T = 213 K.
Figure 3: The structure and orientation of the chromophore 11-cis retinal in the binding pocket of the GPCR protein bovine rhodopsin in its dark-adapted ground state (left), including the experimentally obtained structural constraints (helix 7 of the protein used as z-axis reference; helix only partially displayed).

Similar content being viewed by others

References

  1. Birge, R. R. Nature of the primary photochemical events in rhodopsin and bacteriorhodopsin. Biochim. Biophys. Acta 1016, 293– 327 (1990).

    Article  CAS  Google Scholar 

  2. Krebs, A., Villa, C., Edwards, P. C. & Schertler, G. F. X. Characterisation of an improved two-dimensional p22121 crystal from bovine rhodopsin. J. Mol. Biol. 282, 991–1003 (1998).

    Article  CAS  Google Scholar 

  3. Baldwin, J. M., Schertler, G. F. X. & Unger, V. M. An alpha-carbon template for the transmembrane helices in the rhodopsin family of G-protein-coupled receptors. J. Mol. Biol. 272, 144–164 ( 1997).

    Article  CAS  Google Scholar 

  4. Baldwin, J. M. Structure and function of receptors coupled to G proteins. Curr. Opin. Cell Biol. 6, 180–190 (1994).

    Article  CAS  Google Scholar 

  5. Donnelly, D. & Findlay, J. B. C. Seven-helix receptors: structure and modelling. Curr. Opin. Struct. Biol. 4, 582–589 (1994).

    Article  CAS  Google Scholar 

  6. Herzyk, P. & Hubbard, R. E. Combined biophysical and biochemical information confirms arrangement of transmembrane helices visible from the three-dimensional map of frog rhodopsin. J. Mol. Biol. 281, 741–754 (1998).

    Article  CAS  Google Scholar 

  7. Glaubitz, C., Burnett, I. J., Gröbner, G., Mason, A. J. & Watts, A. Deuterium-MAS NMR spectroscopy on oriented membrane proteins: applications to photointermediates of bacteriorhodopsin. J. Am. Chem. Soc. 121, 5787 ( 1999).

    Article  CAS  Google Scholar 

  8. Ji, T. H., Grossmann, M. & Ji, I. G Protein-coupled receptors. J. Biol. Chem. 273, 17299–17302 ( 1998).

    Article  CAS  Google Scholar 

  9. Altenbach, C., Cai, K., Khorana, H. G. & Hubbell, W. L. Structural features and light-dependent changes in the sequence 306–322 extending from helix VII to the palmitoylation sites in rhodopsin: a site-directed spin-labeling study. Biochemistry 38, 7931– 7937 (1999).

    Article  CAS  Google Scholar 

  10. Tan, Q. et al. Absolute sense of twist of the C12–C13 bond of the retinal chromophore in bovine rhodopsin based on exciton-coupled CD spectra of 11,12-dihydroretinal analogues. Angew. Chem Int. Ed. 36, 2089 –2093 (1997).

    Article  CAS  Google Scholar 

  11. Buss, V., Kolster, K., Terstegen, F. & Vahrenhorst, R. Absolute sense of twist of the C12–C13 bond of the retinal chromophore in rhodopsin-semiempirical and nonempirical calculations of chiroptical data. Angew. Chem Int. Ed. 37, 1893– 1895 (1998).

    Article  CAS  Google Scholar 

  12. Rothschild, K. J., Sanches, R., Hsiao, T. L. & Clark, N. A. A spectroscopic study of rhodopsin alpha-helix orientation. Biophys. J. 31, 53–64 ( 1980).

    Article  CAS  Google Scholar 

  13. Jaeger, S. et al. Chromophore structural changes in rhodopsin form nanoseconds to microseconds following pigment photolysis. Proc. Natl Acad. Sci. USA 94, 8557–8562 ( 1997).

    Article  ADS  Google Scholar 

  14. Shieh, T., Han, M., Sakmar, T. P. & Smith, S. O. The steric trigger in rhodopsin activation. J. Mol. Biol. 269, 373–384 (1997).

    Article  CAS  Google Scholar 

  15. Eilers, M., Reeves, P. J., Ying, W., Khorana, H. G. & Smith, S. O. Magic angle spinning NMR of the protonated retinylidene Schiff base nitrogen in rhodopsin: Expression of 15N-lysine- and 13 C-glycine-labeled opsin in a stable cell line. Proc. Natl Acad. Sci. USA 96, 487– 492 (1999).

    Article  ADS  CAS  Google Scholar 

  16. Feng, X. et al. Direct determination of a molecular torsional angle in the membrane protein rhodopsin by solid-state NMR. J. Am. Chem. Soc. 119, 6853–6857 (1997).

    Article  CAS  Google Scholar 

  17. Verdegem, P. J. E. et al. Retinylidene ligand structure in bovine rhodopsin, metarhodopsin-I, and 10-methylrhodopsin from internuclear distance measurements using 13C-labeling and 1D rotational resonance MAS NMR. Biochemistry 33, 11316–11324 ( 1999).

    Article  Google Scholar 

  18. Feng, X. et al. Determination of a molecular torsional angle in the metarhodopsin-I photointermediate of rhodopsin by double-quantum solid-state NMR. J. Biomol. NMR 16, 1–8 ( 2000).

    Article  CAS  Google Scholar 

  19. Gröbner, G. et al. Photoreceptor rhodopsin: structural and conformational study of its chromophore 11-cis retinal in oriented membranes by deuterium solid state NMR. FEBS Lett. 422, 201– 204 (1998).

    Article  Google Scholar 

  20. Gröbner, G. et al. Macroscopic orientation of natural and model membranes for structural studies. Anal. Biochem. 254, 132–138 (1997).

    Article  Google Scholar 

  21. Smith, S. O. et al. Low-temperature solid-state 13C NMR studies of the retinal chromophore in rhodopsin. Biochemistry 26, 1606–1611 (1987).

    Article  CAS  Google Scholar 

  22. Hudson, B. S. & Birge, R. R. Angular orientation of the retinyl chromophore of bacteriorhodopsin: Reconciliation of 2H NMR and optical measurements. J. Phys. Chem. A 103, 2274–2281 (1999).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank P. Fisher and B. Bonev for help, and J. Lugtenburg for discussion and advice. This work was supported by grants to A.W. (MRC, BBSRC, EU TMR, EU Biotech , Magnex, Varian, Bruker and HEFCE); C.G. is supported by a DFG Emmy Noether Fellowship.

Author information

Author notes

  1. Gerhard Gröbner

    Present address: Department of Chemistry,Biophysical Chemistry, Umeå University, SE-901 87, Umeå, Sweden

  2. Gregory Choi

    Present address: University of Connecticut, Molecular & Cell Biology, LSA 205, 75 North Eagleville Road , U-125, Storrs, Connecticut, 06269-3125, USA

  3. Correspondence and requests for materials should be addressed to A.W. The structural data for the chromophore can be obtained from the authors and will be posted at http://www.bioch.ox.ac.uk/~watts.

Authors and Affiliations

  1. Department of Biochemistry, Biomembrane Structure Unit, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK

    Gerhard Gröbner, Ian J. Burnett, Clemens Glaubitz, Gregory Choi, A. James Mason & Anthony Watts

Authors
  1. Gerhard Gröbner
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ian J. Burnett
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Clemens Glaubitz
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Gregory Choi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. A. James Mason
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Anthony Watts
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Anthony Watts.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gröbner, G., Burnett, I., Glaubitz, C. et al. Observations of light-induced structural changes of retinal within rhodopsin . Nature 405, 810–813 (2000). https://doi.org/10.1038/35015604

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/35015604

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing