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CODA-RET reveals functional selectivity as a result of GPCR heteromerization

Nature Chemical Biology volume 7, pages 624–630 (2011)Cite this article

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Abstract

Here we present a new method that combines protein complementation with resonance energy transfer to study conformational changes in response to activation of a defined G protein–coupled receptor heteromer, and we apply the approach to the putative dopamine D1-D2 receptor heteromer. Remarkably, the potency of the D2 dopamine receptor (D2R) agonist R-(−)-10,11-dihydroxy-N-n-propylnoraporphine (NPA) to change the Gαi conformation via the D2R protomer in the D1-D2 heteromer was enhanced ten-fold relative to its potency in the D2R homomer. In contrast, the potencies of the D2R agonists dopamine and quinpirole were the same in the homomer and heteromer. Thus, we have uncovered a molecular mechanism for functional selectivity in which a drug acts differently at a G protein–coupled receptor (GPCR) protomer depending on the identity of the second protomer participating in the formation of the signaling unit—opening the door to enhancing pharmacological specificity by targeting differences between homomeric and heteromeric signaling.

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Figure 1: D1R coexpression enhances NPA potency at D2R.
Figure 2: Characterization of homomeric receptors in G protein signaling complexes.
Figure 3: Characterization of heteromeric receptors in G protein signaling complexes.
Figure 4: CODA-RET reveals functional selectivity of NPA at the D2R-D1R heteromer.

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Acknowledgements

This work was supported in part by US National Institutes of Health (NIH) grants DA022413 and MH054137 (J.A.J.), GM078319 (N.L.) and NIH TL1 RR024158-04 (H.Y.) as well as by the Lieber Center for Schizophrenia Research and Treatment and EMBO Long-Term and Basque Country fellowships (E.U.).

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Author notes

  1. Eneko Urizar

    Present address: Present address: Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire, Brussels, Belgium.,

  2. Eneko Urizar and Hideaki Yano: These authors contributed equally to this work.

Authors and Affiliations

  1. Center for Molecular Recognition and Departments of Psychiatry and Pharmacology, College of Physicians and Surgeons, Columbia University, New York, New York, USA

    Eneko Urizar, Hideaki Yano, Rachel Kolster & Jonathan A Javitch

  2. Division of Molecular Therapeutics, New York State Psychiatric Institute, New York, New York, USA

    Eneko Urizar & Jonathan A Javitch

  3. Institut National de la Santé et de la Recherche Médicale, Toulouse, France

    Céline Galés

  4. Paul Sabatier University, Toulouse, France

    Céline Galés

  5. Department of Pharmacology and Toxicology, Georgia Health Sciences University, Augusta, Georgia, USA

    Nevin Lambert

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Contributions

E.U., H.Y. and R.K. performed experiments; E.U., H.Y. and J.A.J. planned experiments; J.A.J. supervised the project; C.G. and N.L. provided reagents and discussed the experimental findings and interpretation of results; E.U. and J.A.J. wrote the manuscript.

Corresponding author

Correspondence to Jonathan A Javitch.

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The authors declare no competing financial interests.

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Urizar, E., Yano, H., Kolster, R. et al. CODA-RET reveals functional selectivity as a result of GPCR heteromerization. Nat Chem Biol 7, 624–630 (2011). https://doi.org/10.1038/nchembio.623

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