Delineation of the GPR15 receptor-mediated Gα protein signalling profile in recombinant mammalian cells

Yufang Deng; Ee Von Moo; Claudia Victoria Pérez Almería; Patrick R. Gentry; Line Vedel; Jesper M. Mathiesen; Hans Bräuner-Osborne

doi:10.1111/bcpt.13738

Delineation of the GPR15 receptor-mediated Gα protein signalling profile in recombinant mammalian cells

Yufang Deng, Ee Von Moo, Claudia Victoria Pérez Almería, Patrick R. Gentry, Line Vedel, Jesper M. Mathiesen, Hans Bräuner-Osborne^*

^*Corresponding author for this work

Research output: Contribution to Journal › Article › Academic › peer-review

Abstract

The GPR15 receptor is a G protein-coupled receptor (GPCR), which is activated by an endogenous peptide GPR15L(25–81) and a C-terminal peptide fragment GPR15L(71–81). GPR15 signals through the G_i/o pathway to decrease intracellular cyclic adenosine 3′,5′-monophosphate (cAMP). However, the activation profiles of the GPR15 receptor within G_i/o subtypes have not been examined. Moreover, whether the receptor can also couple to G_s, G_q/11 and G_12/13 is unclear. Here, GPR15L(25–81) and GPR15L(71–81) are used as pharmacological tool compounds to delineate the GPR15 receptor-mediated Gα protein signalling using a G protein activation assay and second messenger assay conducted on living cells. The results show that the GPR15 receptor preferentially couples to G_i/o rather than other pathways in both assays. Within the G_i/o family, the GPR15 receptor activates all the subtypes (G_i1, G_i2, G_i3, G_oA, G_oB and G_z). The E_max and activation rates of G_i1, G_i2, G_i3, G_oA and G_oB are similar, whilst the E_max of G_z is smaller and the activation rate is significantly slower. The potencies of both peptides toward each G_i/o subtype have been determined. Furthermore, the GPR15 receptor signals through G_i/o to inhibit cAMP accumulation, which could be blocked by the application of the G_i/o inhibitor pertussis toxin.

Original language	English
Pages (from-to)	104-113
Number of pages	10
Journal	Basic and Clinical Pharmacology and Toxicology
Volume	131
Issue number	2
Early online date	5 May 2022
DOIs	https://doi.org/10.1111/bcpt.13738
Publication status	Published - Aug 2022

Bibliographical note

Funding Information:
This project was funded by the China Scholarship Council (Grant #201907940002 to Y.D.), the Novo Nordisk Foundation (Grants NNF17OC0028412 and NNF19OC0057730 to H.B.‐O.), the Carlsberg Foundation (Grant CF20‐0248 to H.B.‐O.) and the European Union's Horizon2020 research and innovation program under the Marie Sklodowska‐Curie grant agreement No 846827 (E.V.M.)

Funding Information:
This project was funded by the China Scholarship Council (Grant #201907940002 to Y.D.), the Novo Nordisk Foundation (Grants NNF17OC0028412 and NNF19OC0057730 to H.B.-O.), the Carlsberg Foundation (Grant CF20-0248 to H.B.-O.) and the European Union's Horizon2020 research and innovation program under the Marie Sklodowska-Curie grant agreement No 846827 (E.V.M.)

Publisher Copyright:
© 2022 The Authors. Basic & Clinical Pharmacology & Toxicology published by John Wiley & Sons Ltd on behalf of Nordic Association for the Publication of BCPT (former Nordic Pharmacological Society).

Funding

This project was funded by the China Scholarship Council (Grant #201907940002 to Y.D.), the Novo Nordisk Foundation (Grants NNF17OC0028412 and NNF19OC0057730 to H.B.‐O.), the Carlsberg Foundation (Grant CF20‐0248 to H.B.‐O.) and the European Union's Horizon2020 research and innovation program under the Marie Sklodowska‐Curie grant agreement No 846827 (E.V.M.) This project was funded by the China Scholarship Council (Grant #201907940002 to Y.D.), the Novo Nordisk Foundation (Grants NNF17OC0028412 and NNF19OC0057730 to H.B.-O.), the Carlsberg Foundation (Grant CF20-0248 to H.B.-O.) and the European Union's Horizon2020 research and innovation program under the Marie Sklodowska-Curie grant agreement No 846827 (E.V.M.)

Funders	Funder number
European Union's Horizon2020 research and innovation program
Horizon 2020 Framework Programme
H2020 Marie Skłodowska-Curie Actions	846827
Carlsbergfondet	CF20‐0248
China Scholarship Council	201907940002
Novo Nordisk Fonden	NNF17OC0028412, NNF19OC0057730

Keywords

BRET
GPR15
Gα protein
second messenger
signalling

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1111/bcpt.13738

Persistent URL (handle)

Persistent link

Cite this

@article{123ae55144fd47ff8167d15bad897b87,

title = "Delineation of the GPR15 receptor-mediated Gα protein signalling profile in recombinant mammalian cells",

abstract = "The GPR15 receptor is a G protein-coupled receptor (GPCR), which is activated by an endogenous peptide GPR15L(25–81) and a C-terminal peptide fragment GPR15L(71–81). GPR15 signals through the Gi/o pathway to decrease intracellular cyclic adenosine 3′,5′-monophosphate (cAMP). However, the activation profiles of the GPR15 receptor within Gi/o subtypes have not been examined. Moreover, whether the receptor can also couple to Gs, Gq/11 and G12/13 is unclear. Here, GPR15L(25–81) and GPR15L(71–81) are used as pharmacological tool compounds to delineate the GPR15 receptor-mediated Gα protein signalling using a G protein activation assay and second messenger assay conducted on living cells. The results show that the GPR15 receptor preferentially couples to Gi/o rather than other pathways in both assays. Within the Gi/o family, the GPR15 receptor activates all the subtypes (Gi1, Gi2, Gi3, GoA, GoB and Gz). The Emax and activation rates of Gi1, Gi2, Gi3, GoA and GoB are similar, whilst the Emax of Gz is smaller and the activation rate is significantly slower. The potencies of both peptides toward each Gi/o subtype have been determined. Furthermore, the GPR15 receptor signals through Gi/o to inhibit cAMP accumulation, which could be blocked by the application of the Gi/o inhibitor pertussis toxin.",

keywords = "BRET, GPR15, Gα protein, second messenger, signalling",

author = "Yufang Deng and Moo, {Ee Von} and {P{\'e}rez Almer{\'i}a}, {Claudia Victoria} and Gentry, {Patrick R.} and Line Vedel and Mathiesen, {Jesper M.} and Hans Br{\"a}uner-Osborne",

note = "Funding Information: This project was funded by the China Scholarship Council (Grant #201907940002 to Y.D.), the Novo Nordisk Foundation (Grants NNF17OC0028412 and NNF19OC0057730 to H.B.‐O.), the Carlsberg Foundation (Grant CF20‐0248 to H.B.‐O.) and the European Union's Horizon2020 research and innovation program under the Marie Sklodowska‐Curie grant agreement No 846827 (E.V.M.) Funding Information: This project was funded by the China Scholarship Council (Grant #201907940002 to Y.D.), the Novo Nordisk Foundation (Grants NNF17OC0028412 and NNF19OC0057730 to H.B.-O.), the Carlsberg Foundation (Grant CF20-0248 to H.B.-O.) and the European Union's Horizon2020 research and innovation program under the Marie Sklodowska-Curie grant agreement No 846827 (E.V.M.) Publisher Copyright: {\textcopyright} 2022 The Authors. Basic & Clinical Pharmacology & Toxicology published by John Wiley & Sons Ltd on behalf of Nordic Association for the Publication of BCPT (former Nordic Pharmacological Society).",

year = "2022",

month = aug,

doi = "10.1111/bcpt.13738",

language = "English",

volume = "131",

pages = "104--113",

journal = "Basic and Clinical Pharmacology and Toxicology",

issn = "1742-7835",

publisher = "Wiley-Blackwell",

number = "2",

}

TY - JOUR

T1 - Delineation of the GPR15 receptor-mediated Gα protein signalling profile in recombinant mammalian cells

AU - Deng, Yufang

AU - Moo, Ee Von

AU - Pérez Almería, Claudia Victoria

AU - Gentry, Patrick R.

AU - Vedel, Line

AU - Mathiesen, Jesper M.

AU - Bräuner-Osborne, Hans

N1 - Funding Information: This project was funded by the China Scholarship Council (Grant #201907940002 to Y.D.), the Novo Nordisk Foundation (Grants NNF17OC0028412 and NNF19OC0057730 to H.B.‐O.), the Carlsberg Foundation (Grant CF20‐0248 to H.B.‐O.) and the European Union's Horizon2020 research and innovation program under the Marie Sklodowska‐Curie grant agreement No 846827 (E.V.M.) Funding Information: This project was funded by the China Scholarship Council (Grant #201907940002 to Y.D.), the Novo Nordisk Foundation (Grants NNF17OC0028412 and NNF19OC0057730 to H.B.-O.), the Carlsberg Foundation (Grant CF20-0248 to H.B.-O.) and the European Union's Horizon2020 research and innovation program under the Marie Sklodowska-Curie grant agreement No 846827 (E.V.M.) Publisher Copyright: © 2022 The Authors. Basic & Clinical Pharmacology & Toxicology published by John Wiley & Sons Ltd on behalf of Nordic Association for the Publication of BCPT (former Nordic Pharmacological Society).

PY - 2022/8

Y1 - 2022/8

N2 - The GPR15 receptor is a G protein-coupled receptor (GPCR), which is activated by an endogenous peptide GPR15L(25–81) and a C-terminal peptide fragment GPR15L(71–81). GPR15 signals through the Gi/o pathway to decrease intracellular cyclic adenosine 3′,5′-monophosphate (cAMP). However, the activation profiles of the GPR15 receptor within Gi/o subtypes have not been examined. Moreover, whether the receptor can also couple to Gs, Gq/11 and G12/13 is unclear. Here, GPR15L(25–81) and GPR15L(71–81) are used as pharmacological tool compounds to delineate the GPR15 receptor-mediated Gα protein signalling using a G protein activation assay and second messenger assay conducted on living cells. The results show that the GPR15 receptor preferentially couples to Gi/o rather than other pathways in both assays. Within the Gi/o family, the GPR15 receptor activates all the subtypes (Gi1, Gi2, Gi3, GoA, GoB and Gz). The Emax and activation rates of Gi1, Gi2, Gi3, GoA and GoB are similar, whilst the Emax of Gz is smaller and the activation rate is significantly slower. The potencies of both peptides toward each Gi/o subtype have been determined. Furthermore, the GPR15 receptor signals through Gi/o to inhibit cAMP accumulation, which could be blocked by the application of the Gi/o inhibitor pertussis toxin.

AB - The GPR15 receptor is a G protein-coupled receptor (GPCR), which is activated by an endogenous peptide GPR15L(25–81) and a C-terminal peptide fragment GPR15L(71–81). GPR15 signals through the Gi/o pathway to decrease intracellular cyclic adenosine 3′,5′-monophosphate (cAMP). However, the activation profiles of the GPR15 receptor within Gi/o subtypes have not been examined. Moreover, whether the receptor can also couple to Gs, Gq/11 and G12/13 is unclear. Here, GPR15L(25–81) and GPR15L(71–81) are used as pharmacological tool compounds to delineate the GPR15 receptor-mediated Gα protein signalling using a G protein activation assay and second messenger assay conducted on living cells. The results show that the GPR15 receptor preferentially couples to Gi/o rather than other pathways in both assays. Within the Gi/o family, the GPR15 receptor activates all the subtypes (Gi1, Gi2, Gi3, GoA, GoB and Gz). The Emax and activation rates of Gi1, Gi2, Gi3, GoA and GoB are similar, whilst the Emax of Gz is smaller and the activation rate is significantly slower. The potencies of both peptides toward each Gi/o subtype have been determined. Furthermore, the GPR15 receptor signals through Gi/o to inhibit cAMP accumulation, which could be blocked by the application of the Gi/o inhibitor pertussis toxin.

KW - BRET

KW - GPR15

KW - Gα protein

KW - second messenger

KW - signalling

UR - http://www.scopus.com/inward/record.url?scp=85130999840&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85130999840&partnerID=8YFLogxK

U2 - 10.1111/bcpt.13738

DO - 10.1111/bcpt.13738

M3 - Article

C2 - 35510660

AN - SCOPUS:85130999840

SN - 1742-7835

VL - 131

SP - 104

EP - 113

JO - Basic and Clinical Pharmacology and Toxicology

JF - Basic and Clinical Pharmacology and Toxicology

IS - 2

ER -

Delineation of the GPR15 receptor-mediated Gα protein signalling profile in recombinant mammalian cells

Abstract

Bibliographical note

Funding

Keywords

UN SDGs

Access to Document

Persistent URL (handle)

Other files and links

Fingerprint

Cite this