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Oligomerization of G-protein-coupled receptors shown by selective co-immunoprecipitation

Oligomerization of G-protein-coupled receptors shown by selective co-immunoprecipitation
Oligomerization of G-protein-coupled receptors shown by selective co-immunoprecipitation
Recent studies have shown that G-protein-coupled receptors (GPCRs) can assemble as high molecular weight homo- and hetero-oligomeric complexes. This can result in altered receptor-ligand binding, signaling, or intracellular trafficking. We have co-transfected HEK-293 cells with differentially epitope-tagged GPCRs from different subfamilies and determined whether oligomeric complexes were formed by co-immunoprecipitation and immunoblot analysis. This gave the surprising result that the 5HT(1A) receptor was capable of forming hetero-oligomers with all GPCRs tested including the 5HT(1B), 5HT(1D), EDG(1), EDG(3), GPR(26), and GABA(B2) receptors. The testing of other GPCR combinations showed similar results with hetero-oligomer formation occurring for the 5HT(1D) with the 5HT(1B) and EDG(1) receptor. Control studies showed that these complexes were present in co-transfected cells before the time of lysis and that the hetero-oligomers were comprised of GPCRs at discrete stoichiometries. These findings suggest that GPCRs have a natural tendency to form oligomers when co-transfected into cells. Future studies should therefore investigate the presence and physiological role of GPCR hetero-oligomers in cells in which they are endogenously expressed.
Cell Line, GTP-Binding Proteins, Humans, Immunoblotting, Kidney, Mutagenesis, Site-Directed, Protein Subunits, Proto-Oncogene Proteins c-myc, Receptor, Serotonin, 5-HT1B, 5-HT1D, Receptors, Cell Surface, 5-HT1, Recombinant Proteins, Restriction Mapping, Transfection
1083-351X
15482-15485
Salim, K
3f7c5436-8bbd-413c-9890-71aa7d87a07b
Fenton, TR
087260ba-f6a1-405a-85df-099d05810a84
Bacha, J
bd721bc6-3630-4fb1-b0a2-a93d589cc4c1
Urien-Rodriguez, H
17f1fc95-310f-4336-bca5-27e6a7853a3a
Bonnert, T
b3bb5fc9-634f-425f-b1ff-271c8d2f05a7
Skynner, HA
bd1fd777-4732-4e6a-aeb4-a0099e55bc0d
Watts, E
3a7836fd-a182-4e65-876a-f0f3815cbe8e
Kerby, J
0806187d-9886-4d29-b6c5-7b8d55f1dc10
Heald, A
c686c304-f42d-4c61-abda-200764278f77
Beer, M
e907ff45-441b-466a-9e2f-4da93fad1e94
McAllister, G
0a12e929-b90d-4376-b01a-ec303ce11fcc
Guest, PC
39cb8c38-ca95-4cc7-b8fd-091bf147c14e
Salim, K
3f7c5436-8bbd-413c-9890-71aa7d87a07b
Fenton, TR
087260ba-f6a1-405a-85df-099d05810a84
Bacha, J
bd721bc6-3630-4fb1-b0a2-a93d589cc4c1
Urien-Rodriguez, H
17f1fc95-310f-4336-bca5-27e6a7853a3a
Bonnert, T
b3bb5fc9-634f-425f-b1ff-271c8d2f05a7
Skynner, HA
bd1fd777-4732-4e6a-aeb4-a0099e55bc0d
Watts, E
3a7836fd-a182-4e65-876a-f0f3815cbe8e
Kerby, J
0806187d-9886-4d29-b6c5-7b8d55f1dc10
Heald, A
c686c304-f42d-4c61-abda-200764278f77
Beer, M
e907ff45-441b-466a-9e2f-4da93fad1e94
McAllister, G
0a12e929-b90d-4376-b01a-ec303ce11fcc
Guest, PC
39cb8c38-ca95-4cc7-b8fd-091bf147c14e

Salim, K, Fenton, TR, Bacha, J, Urien-Rodriguez, H, Bonnert, T, Skynner, HA, Watts, E, Kerby, J, Heald, A, Beer, M, McAllister, G and Guest, PC (2002) Oligomerization of G-protein-coupled receptors shown by selective co-immunoprecipitation. Journal of Biological Chemistry, 277 (18), 15482-15485. (doi:10.1074/jbc.M201539200).

Record type: Article

Abstract

Recent studies have shown that G-protein-coupled receptors (GPCRs) can assemble as high molecular weight homo- and hetero-oligomeric complexes. This can result in altered receptor-ligand binding, signaling, or intracellular trafficking. We have co-transfected HEK-293 cells with differentially epitope-tagged GPCRs from different subfamilies and determined whether oligomeric complexes were formed by co-immunoprecipitation and immunoblot analysis. This gave the surprising result that the 5HT(1A) receptor was capable of forming hetero-oligomers with all GPCRs tested including the 5HT(1B), 5HT(1D), EDG(1), EDG(3), GPR(26), and GABA(B2) receptors. The testing of other GPCR combinations showed similar results with hetero-oligomer formation occurring for the 5HT(1D) with the 5HT(1B) and EDG(1) receptor. Control studies showed that these complexes were present in co-transfected cells before the time of lysis and that the hetero-oligomers were comprised of GPCRs at discrete stoichiometries. These findings suggest that GPCRs have a natural tendency to form oligomers when co-transfected into cells. Future studies should therefore investigate the presence and physiological role of GPCR hetero-oligomers in cells in which they are endogenously expressed.

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More information

e-pub ahead of print date: 19 February 2002
Published date: 3 May 2002
Keywords: Cell Line, GTP-Binding Proteins, Humans, Immunoblotting, Kidney, Mutagenesis, Site-Directed, Protein Subunits, Proto-Oncogene Proteins c-myc, Receptor, Serotonin, 5-HT1B, 5-HT1D, Receptors, Cell Surface, 5-HT1, Recombinant Proteins, Restriction Mapping, Transfection

Identifiers

Local EPrints ID: 453956
URI: http://eprints.soton.ac.uk/id/eprint/453956
ISSN: 1083-351X
PURE UUID: 81c62e67-b79e-4ad8-b7bc-03760785514d
ORCID for TR Fenton: ORCID iD orcid.org/0000-0002-4737-8233

Catalogue record

Date deposited: 26 Jan 2022 17:48
Last modified: 17 Mar 2024 04:11

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Contributors

Author: K Salim
Author: TR Fenton ORCID iD
Author: J Bacha
Author: H Urien-Rodriguez
Author: T Bonnert
Author: HA Skynner
Author: E Watts
Author: J Kerby
Author: A Heald
Author: M Beer
Author: G McAllister
Author: PC Guest

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