Functional homomers and heteromers of dopamine D2L and D3 receptors co-exist at the cell surface

Pou, C., Mannoury la Cour, C., Stoddart, L.A., Millan, M.J. and Milligan, G. (2012) Functional homomers and heteromers of dopamine D2L and D3 receptors co-exist at the cell surface. Journal of Biological Chemistry, 287(12), pp. 8864-8878. (doi: 10.1074/jbc.M111.326678)

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Publisher's URL: http://dx.doi.org/10.1074/jbc.M111.326678

Abstract

Human dopamine D<sub>2long</sub> and D<sub>3</sub> receptors were modified by N-terminal addition of SNAP or CLIP forms of C<sup>6</sup>-alkylguanine-DNA-alkyltransferase plus a peptide epitope tag. Cells able to express each of these four constructs only upon addition of an antibiotic were established and used to confirm regulated and inducible control of expression, the specificity of SNAP and CLIP tag covalent labeling reagents, and based on homogenous time-resolved fluorescence resonance energy transfer, the presence of cell surface D<sub>2long</sub> and D<sub>3</sub> receptor homomers. Following constitutive expression of reciprocal constructs, potentially capable of forming and reporting the presence of cell surface D<sub>2long</sub>-D<sub>3</sub> heteromers, individual clones were assessed for levels of expression of the constitutively expressed protomer. This was unaffected by induction of the partner protomer and the level of expression of the partner required to generate detectable cell surface D<sub>2long</sub>–D<sub>3</sub> heteromers was defined. Such homomers and heteromers were found to co-exist and using a reconstitution of function approach both homomers and heteromers of D<sub>2long</sub> and D<sub>3</sub> receptors were shown to be functional, potentially via trans-activation of associated G protein. These studies demonstrate the ability of dopamine D<sub>2long</sub> and D<sub>3</sub> receptors to form both homomers and heteromers, and show that in cells expressing each subtype a complex mixture of homomers and heteromers co-exists at steady state. These data are of potential importance both to disorders in which D<sub>2long</sub> and D<sub>3</sub> receptors are implicated, like schizophrenia and Parkinson disease, and also to drugs exerting their actions via these sites.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Millan, Dr Mark and Milligan, Professor Graeme and Pou, Dr Chantevy
Authors: Pou, C., Mannoury la Cour, C., Stoddart, L.A., Millan, M.J., and Milligan, G.
College/School:College of Medical Veterinary and Life Sciences > Institute of Molecular Cell and Systems Biology
College of Medical Veterinary and Life Sciences > School of Psychology & Neuroscience
Journal Name:Journal of Biological Chemistry
Journal Abbr.:J Biol Chem.
ISSN:0021-9258
ISSN (Online):1083-351X
Published Online:16 March 2012

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
G0900050