G Protein-coupled Receptor 30 (GPR30) Forms a Plasma Membrane Complex With Membrane-associated Guanylate Kinases (MAGUKs) and AKAP5 That Constitutively Inhibits cAMP Production.
(2014) In Journal of Biological Chemistry 289(32). p.22117-22127- Abstract
- GPR30, or G protein-coupled estrogen receptor (GPER), is a GPCR reported to bind 17β-estradiol (E2), couple to the G proteins Gs and Gi/o, and mediate non-genomic estrogenic responses. However, controversies exist regarding the receptor pharmacological profile, effector coupling, and subcellular localization. We addressed the role of the type I PSD-95/Discs-large/ZO-1 homology (PDZ) motif at the receptor C-terminus in receptor trafficking and coupling to cAMP production in HEK293 cells and CHO cells ectopically expressing the receptor, and in MDCK cells expressing native receptor. GPR30 was localized both intracellularly and in the plasma membrane and subject to limited basal endocytosis. E2 and G-1, reported GPR30 agonists, neither... (More)
- GPR30, or G protein-coupled estrogen receptor (GPER), is a GPCR reported to bind 17β-estradiol (E2), couple to the G proteins Gs and Gi/o, and mediate non-genomic estrogenic responses. However, controversies exist regarding the receptor pharmacological profile, effector coupling, and subcellular localization. We addressed the role of the type I PSD-95/Discs-large/ZO-1 homology (PDZ) motif at the receptor C-terminus in receptor trafficking and coupling to cAMP production in HEK293 cells and CHO cells ectopically expressing the receptor, and in MDCK cells expressing native receptor. GPR30 was localized both intracellularly and in the plasma membrane and subject to limited basal endocytosis. E2 and G-1, reported GPR30 agonists, neither stimulated nor inhibited cAMP production through GPR30, nor influenced receptor localization. Instead, GPR30 constitutively inhibited cAMP production stimulated by a heterologous agonist independently of Gi/o. Moreover, siRNA knockdown of native GPR30 increased cAMP production. Deletion of the receptor PDZ motif interfered with inhibition of cAMP production and increased basal receptor endocytosis. GPR30 interacted with membrane-associated guanylate kinases (MAGUKs), including SAP97 and PSD-95, and A-kinase anchoring protein (AKAP) 5 in the plasma membrane in a PDZ-dependent manner. Knockdown of AKAP5 or St-Ht31 treatment, to disrupt AKAP interaction with protein kinase A (PKA) RIIβ regulatory subunit, decreased inhibition of cAMP production, and St-Ht31 increased basal receptor endocytosis. Thus, GPR30 forms a plasma membrane complex with a MAGUK and AKAP5, which constitutively attenuates cAMP production in response to heterologous agonists independently of Gi/o and retains receptors in the plasma membrane. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/4526842
- author
- Broselid, Stefan LU ; Berg, Kelly A ; Chavera, Teresa A ; Kahn, Robin LU ; Clarke, William P ; Olde, Björn LU and Leeb-Lundberg, Fredrik LU
- organization
- publishing date
- 2014
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of Biological Chemistry
- volume
- 289
- issue
- 32
- pages
- 22117 - 22127
- publisher
- American Society for Biochemistry and Molecular Biology
- external identifiers
-
- pmid:24962572
- wos:000340593500027
- scopus:84905844102
- pmid:24962572
- ISSN
- 1083-351X
- DOI
- 10.1074/jbc.M114.566893
- language
- English
- LU publication?
- yes
- id
- 2822880f-f30d-4b78-a480-1a6e2d66fc07 (old id 4526842)
- alternative location
- http://www.ncbi.nlm.nih.gov/pubmed/24962572?dopt=Abstract
- date added to LUP
- 2016-04-01 10:39:29
- date last changed
- 2025-04-04 14:40:31
@article{2822880f-f30d-4b78-a480-1a6e2d66fc07,
abstract = {{GPR30, or G protein-coupled estrogen receptor (GPER), is a GPCR reported to bind 17β-estradiol (E2), couple to the G proteins Gs and Gi/o, and mediate non-genomic estrogenic responses. However, controversies exist regarding the receptor pharmacological profile, effector coupling, and subcellular localization. We addressed the role of the type I PSD-95/Discs-large/ZO-1 homology (PDZ) motif at the receptor C-terminus in receptor trafficking and coupling to cAMP production in HEK293 cells and CHO cells ectopically expressing the receptor, and in MDCK cells expressing native receptor. GPR30 was localized both intracellularly and in the plasma membrane and subject to limited basal endocytosis. E2 and G-1, reported GPR30 agonists, neither stimulated nor inhibited cAMP production through GPR30, nor influenced receptor localization. Instead, GPR30 constitutively inhibited cAMP production stimulated by a heterologous agonist independently of Gi/o. Moreover, siRNA knockdown of native GPR30 increased cAMP production. Deletion of the receptor PDZ motif interfered with inhibition of cAMP production and increased basal receptor endocytosis. GPR30 interacted with membrane-associated guanylate kinases (MAGUKs), including SAP97 and PSD-95, and A-kinase anchoring protein (AKAP) 5 in the plasma membrane in a PDZ-dependent manner. Knockdown of AKAP5 or St-Ht31 treatment, to disrupt AKAP interaction with protein kinase A (PKA) RIIβ regulatory subunit, decreased inhibition of cAMP production, and St-Ht31 increased basal receptor endocytosis. Thus, GPR30 forms a plasma membrane complex with a MAGUK and AKAP5, which constitutively attenuates cAMP production in response to heterologous agonists independently of Gi/o and retains receptors in the plasma membrane.}},
author = {{Broselid, Stefan and Berg, Kelly A and Chavera, Teresa A and Kahn, Robin and Clarke, William P and Olde, Björn and Leeb-Lundberg, Fredrik}},
issn = {{1083-351X}},
language = {{eng}},
number = {{32}},
pages = {{22117--22127}},
publisher = {{American Society for Biochemistry and Molecular Biology}},
series = {{Journal of Biological Chemistry}},
title = {{G Protein-coupled Receptor 30 (GPR30) Forms a Plasma Membrane Complex With Membrane-associated Guanylate Kinases (MAGUKs) and AKAP5 That Constitutively Inhibits cAMP Production.}},
url = {{http://dx.doi.org/10.1074/jbc.M114.566893}},
doi = {{10.1074/jbc.M114.566893}},
volume = {{289}},
year = {{2014}},
}