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Zinc Is Involved in Depression by Modulating G Protein-Coupled Receptor Heterodimerization

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Abstract

5-Hydroxytryptamine 1A receptor and galanin receptor 1 belong to the G protein-coupled receptors superfamily, and they have been described to heterodimerize triggering an anomalous physiological state that would underlie depression. Zinc supplementation has been widely reported to improve treatment against major depressive disorder. Our work has focused on the study and characterization of these receptors and its relationships with zinc both under purified conditions and in cell culture. To this aim, we have designed a strategy to purify the receptors in a conformationally active state. We have used receptors tagged with the monoclonal Rho-1D4 antibody and employed ligand-assisted purification in order to successfully purify both receptors in a properly folded and active state. The interaction between both purified receptors has been analyzed by surface plasmon resonance in order to determine the kinetics of dimerization. Zinc effect on heteromer has also been tested using the same methodology but exposing the 5-hydroxytryptamine 1A receptor to zinc before the binding experiment. These results, combined with Förster resonance energy transfer (FRET) measurements, in the absence and presence of zinc, suggest that this ion is capable of disrupting this interaction. Moreover, molecular modeling suggests that there is a coincidence between zinc-binding sites and heterodimerization interfaces for the serotonin receptor. Our results establish a rational explanation for the role of zinc in the molecular processes associated with receptor-receptor interactions and its relationship with depression, in agreement with previously reported evidence for the positive effects of zinc in depression treatment, and the involvement of our target dimer in the same disease.

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Acknowledgments

We thank Dr. Dasiel Borroto-Escuela for providing pECFP-5-HT1A and pEYFP-GalR1 constructs. This work has been supported by grants from Fundació la Marató de TV3 (090131-01) and AGAUR (2009 SGR 1402) to PG. MT is the recipient of a predoctoral fellowship FPI-UPC from Universitat Politècnica de Catalunya.

Conflict of Interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Departament d’Enginyeria Química, Grup de Biotecnologia Molecular i Industrial, Centre de Biotecnologia Molecular, Universitat Politècnica de Catalunya, Edifici Gaia, Rambla de Sant Nebridi 22, 08222, Terrassa, Catalonia, Spain

    Mercè Tena-Campos, Eva Ramon & Pere Garriga

  2. Departament d’Enginyeria Química, Centre de Biotecnologia Molecular, Universitat Politècnica de Catalunya, ETSEIB, Avda. Diagonal 647, 08028, Barcelona, Catalonia, Spain

    Cecylia S. Lupala & Juan J. Pérez

  3. Department of Neurosciences, Biochemistry Group, University of Oldenburg, Carl-von-Ossietzky-Str. 9-11, 26129, Oldenburg, Germany

    Karl-W. Koch

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  1. Mercè Tena-Campos
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  2. Eva Ramon
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  3. Cecylia S. Lupala
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  4. Juan J. Pérez
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  5. Karl-W. Koch
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  6. Pere Garriga
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Correspondence to Pere Garriga.

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Tena-Campos, M., Ramon, E., Lupala, C.S. et al. Zinc Is Involved in Depression by Modulating G Protein-Coupled Receptor Heterodimerization. Mol Neurobiol 53, 2003–2015 (2016). https://doi.org/10.1007/s12035-015-9153-y

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