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 - Almería, Claudia Victoria Pérez

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.)

PY - 2022

Y1 - 2022

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

U2 - 10.1111/bcpt.13738

DO - 10.1111/bcpt.13738

M3 - Journal article

C2 - 35510660

AN - SCOPUS:85130999840

VL - 131

SP - 104

EP - 113

JO - Basic and Clinical Pharmacology and Toxicology

JF - Basic and Clinical Pharmacology and Toxicology

SN - 1742-7835

IS - 2

ER -