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Sequential Co-immunoprecipitation and Immunoblot Approach to Determine Oligomerisation of G-Protein-Coupled Receptors

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Proteomic Methods in Neuropsychiatric Research

Part of the book series: Advances in Experimental Medicine and Biology ((PMISB,volume 974))

Abstract

G-protein-coupled receptors (GPCRs) play a major role in psychiatric disorders and are the targets of several current therapeutic approaches in this field. A number of studies have now shown that GPCRs can assemble as high molecular weight homo- and hetero-oligomers, which could affect ligand binding, intracellular signalling or trafficking. This information could be critical in design of new drugs to treat neurological and psychiatric disorders. This chapter describes a sequential co-immunoprecipitation and immunoblot protocol for determining oligomerisation of the 5-hydroxytryptamine (HT)1A receptor with other GPCRs in co-transfected HEK-293 cells.

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References

  1. Schonenbach NS, Hussain S, O'Malley MA (2015) Structure and function of G protein-coupled receptor oligomers: implications for drug discovery. Wiley Interdiscip Rev Nanomed Nanobiotechnol 7:408–427

    Article  CAS  PubMed  Google Scholar 

  2. Komatsu H (2015) Novel therapeutic GPCRs for psychiatric disorders. Int J Mol Sci 16:14109–14121

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Franco R, Martínez-Pinilla E, Lanciego JL, Navarro G (2016) Basic pharmacological and structural evidence for class A G-protein-coupled receptor heteromerization. Front Pharmacol 7:76. doi:10.3389/fphar.2016.00076

    Article  PubMed  PubMed Central  Google Scholar 

  4. Salim K, Fenton T, Bacha J, Urien-Rodriguez H, Bonnert T, Skynner HA et al (2002) Oligomerization of G-protein-coupled receptors shown by selective co-immunoprecipitation. J Biol Chem 277:15482–15485

    Article  CAS  PubMed  Google Scholar 

  5. Nelson RM, Long GL (1989) A general method of site-specific mutagenesis using a modification of the Thermus aquaticus polymerase chain reaction. Anal Biochem 180:147–151

    Article  CAS  PubMed  Google Scholar 

  6. Baldi L, Muller N, Picasso S, Jacquet R, Girard P, Thanh HP et al (2005) Transient gene expression in suspension HEK-293 cells: application to large-scale protein production. Biotechnol Prog 21:148–153

    Article  CAS  PubMed  Google Scholar 

  7. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  PubMed  Google Scholar 

  8. Fink K, Zentner J, Gothert M (1995) Subclassification of presynaptic 5-HT autoreceptors in the human cerebral cortex as 5-HT1D receptors. Naunyn Schmiedebergs Arch Pharmacol 352:451–454

    CAS  PubMed  Google Scholar 

  9. Bonaventure P, Voorn P, Luyten WH, Jurzak M, Schotte A, Leysen JE (1998) Detailed mapping of serotonin 5-HT1B and 5-HT1D receptor messenger RNA and ligand binding sites in guinea-pig brain and trigeminal ganglion: clues for function. Neuroscience 82:469–484

    Article  CAS  PubMed  Google Scholar 

  10. Barnes NM, Sharp T (1999) A review of central 5-HT receptors and their function. Neuropharmacology 38:1083–1152

    Article  CAS  PubMed  Google Scholar 

  11. Xie Z, Lee SP, O’Dowd BF, George SR (1999) Serotonin 5-HT1B and 5-HT1D receptors form homodimers when expressed alone and heterodimers when co-expressed. FEBS Lett 456:63–67

    Google Scholar 

  12. Kleinau G, Müller A, Biebermann H (2016) Oligomerization of GPCRs involved in endocrine regulation. J Mol Endocrinol 57:R59–R80

    Article  CAS  PubMed  Google Scholar 

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Correspondence to Paul C. Guest .

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Guest, P.C. (2017). Sequential Co-immunoprecipitation and Immunoblot Approach to Determine Oligomerisation of G-Protein-Coupled Receptors. In: Guest, P. (eds) Proteomic Methods in Neuropsychiatric Research. Advances in Experimental Medicine and Biology(), vol 974. Springer, Cham. https://doi.org/10.1007/978-3-319-52479-5_20

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