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.

  • Correspondence
  • Published:

Experimental challenge to a 'rigorous' BRET analysis of GPCR oligomerization

Nature Methods volume 4, pages 599–600 (2007)Cite this article

To the editor:

James et al.1 describes a new framework for 'rigorous' assessment of bioluminescence resonance energy transfer (BRET) experiments. Although the development of new approaches for BRET analysis is commendable, part of the authors' methodological approach falls short of being 'rigorous'. Specifically, they devise two experimental designs, 'type-1' and 'type-2', to distinguish whether the BRET signal coming from tagged plasma membrane receptors results from constitutive dimerization or random interaction at the plasma membrane. Some of the pitfalls of their type-1 and type-2 experiments have already been discussed elsewhere2. Here we focus on the type-2 experiments, and report experimental data to refute some of the results and conclusions presented in James et al.1.

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

Relevant articles

Open Access articles citing this article.

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: 'Type 2 experiments' for β2AR, D1 and D2-dopamine receptors.

References

  1. James, J.R., Oliveira, M.I., Carmo, A.M., Iaboni, A. & Davis, S.J. Nat. Methods 3, 1001–1006 (2006).

    Article  CAS  Google Scholar 

  2. Bouvier, M., Heveker, N., Jockers, R., Marullo, S. & Milligan, G. Nat. Methods 4, 3–4 (2007).

    Article  CAS  Google Scholar 

  3. Mercier, J.F., Salahpour, A., Angers, S., Breit, A. & Bouvier, M. J. Biol. Chem. 277, 44925–44931 (2002).

    Article  CAS  Google Scholar 

  4. Breit, A., Lagacé, M. & Bouvier, M. J. Biol. Chem. 279, 28756–28765 (2004).

  5. Wang, D., Sun, X., Bohn, L.M. & Sadee, W. Mol. Pharmacol. 67, 2173–2184 (2005).

    Article  CAS  Google Scholar 

  6. Hamdan, F.F., Audet, M., Garneau, P., Pelletier, J. & Bouvier M. J. Biomol. Screen. 5, 463–475 (2005).

    Article  Google Scholar 

Download references

Acknowledgements

We thank M. Caron for support and critical reading of the manuscript, and A. Ramsey for critical reading of the manuscript. Supported by a fellowship from Canadian Institutes of Health Research (A.S.) and by a fellowship from Fondation pour la Recherche Médicale, France (B.M.).

Author information

Authors and Affiliations

  1. Department of Cell Biology, Duke University Medical Center, Durham, 27710, North Carolina, USA

    Ali Salahpour & Bernard Masri

Authors
  1. Ali Salahpour
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bernard Masri
    View author publications

    You can also search for this author in PubMed Google Scholar

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figure 1, Supplementary Tables 1–2, Supplementary Methods (PDF 224 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Salahpour, A., Masri, B. Experimental challenge to a 'rigorous' BRET analysis of GPCR oligomerization. Nat Methods 4, 599–600 (2007). https://doi.org/10.1038/nmeth0807-599

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1038/nmeth0807-599

This article is cited by

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