Human GIP(3-30)NH inhibits G protein-dependent as well as G protein-independent signaling and is selective for the GIP receptor with high-affinity binding to primate but not rodent GIP receptors

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Human GIP(3-30)NH inhibits G protein-dependent as well as G protein-independent signaling and is selective for the GIP receptor with high-affinity binding to primate but not rodent GIP receptors. / Gabe, Maria Buur Nordskov; Sparre-Ulrich, Alexander Hovard; Pedersen, Mie Fabricius; Gasbjerg, Lærke Smidt; Inoue, Asuka; Bräuner-Osborne, Hans; Hartmann, Bolette; Rosenkilde, Mette Marie.

In: Biochemical Pharmacology, Vol. 150, 31.01.2018, p. 97-107.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Gabe, MBN, Sparre-Ulrich, AH, Pedersen, MF, Gasbjerg, LS, Inoue, A, Bräuner-Osborne, H, Hartmann, B & Rosenkilde, MM 2018, 'Human GIP(3-30)NH inhibits G protein-dependent as well as G protein-independent signaling and is selective for the GIP receptor with high-affinity binding to primate but not rodent GIP receptors', Biochemical Pharmacology, vol. 150, pp. 97-107. https://doi.org/10.1016/j.bcp.2018.01.040

APA

Gabe, M. B. N., Sparre-Ulrich, A. H., Pedersen, M. F., Gasbjerg, L. S., Inoue, A., Bräuner-Osborne, H., ... Rosenkilde, M. M. (2018). Human GIP(3-30)NH inhibits G protein-dependent as well as G protein-independent signaling and is selective for the GIP receptor with high-affinity binding to primate but not rodent GIP receptors. Biochemical Pharmacology, 150, 97-107. https://doi.org/10.1016/j.bcp.2018.01.040

Vancouver

Gabe MBN, Sparre-Ulrich AH, Pedersen MF, Gasbjerg LS, Inoue A, Bräuner-Osborne H et al. Human GIP(3-30)NH inhibits G protein-dependent as well as G protein-independent signaling and is selective for the GIP receptor with high-affinity binding to primate but not rodent GIP receptors. Biochemical Pharmacology. 2018 Jan 31;150:97-107. https://doi.org/10.1016/j.bcp.2018.01.040

Author

Gabe, Maria Buur Nordskov ; Sparre-Ulrich, Alexander Hovard ; Pedersen, Mie Fabricius ; Gasbjerg, Lærke Smidt ; Inoue, Asuka ; Bräuner-Osborne, Hans ; Hartmann, Bolette ; Rosenkilde, Mette Marie. / Human GIP(3-30)NH inhibits G protein-dependent as well as G protein-independent signaling and is selective for the GIP receptor with high-affinity binding to primate but not rodent GIP receptors. In: Biochemical Pharmacology. 2018 ; Vol. 150. pp. 97-107.

Bibtex

@article{eee0b4d878fa425083c82f32e25b0693,
title = "Human GIP(3-30)NH inhibits G protein-dependent as well as G protein-independent signaling and is selective for the GIP receptor with high-affinity binding to primate but not rodent GIP receptors",
abstract = "GIP(3-30)NH2is a high affinity antagonist of the GIP receptor (GIPR) in humans inhibiting insulin secretion via G protein-dependent pathways. However, its ability to inhibit G protein-independent signaling is unknown. Here we determine its action on arrestin-recruitment and receptor internalization in recombinant cells. As GIP is adipogenic, we evaluate the inhibitory actions of GIP(3-30)NH2in human adipocytes. Finally, we determine the receptor selectivity of GIP(3-30)NH2among other human and animal GPCRs. cAMP accumulation and β-arrestin 1 and 2 recruitment were studied in transiently transfected HEK293 cells and real-time internalization in transiently transfected HEK293A and in HEK293A β-arrestin 1 and 2 knockout cells. Furthermore, human subcutaneous adipocytes were assessed for cAMP accumulation following ligand stimulation. Competition binding was examined in transiently transfected COS-7 cells using human125I-GIP(3-30)NH2. The selectivity of human GIP(3-30)NH2was examined by testing for agonistic and antagonistic properties on 62 human GPCRs. Human GIP(3-30)NH2inhibited GIP(1-42)-induced cAMP and β-arrestin 1 and 2 recruitment on the human GIPR and Schild plot analysis showed competitive antagonism with a pA2and Hill slope of 16.8 nM and 1.11 ± 0.02 in cAMP, 10.6 nM and 1.15 ± 0.05 in β-arrestin 1 recruitment, and 10.2 nM and 1.06 ± 0.05 in β-arrestin 2 recruitment. Efficient internalization of the GIPR was dependent on the presence of either β-arrestin 1 or 2. Moreover, GIP(3-30)NH2inhibited GIP(1-42)-induced internalization in a concentration-dependent manner and notably also inhibited GIP-mediated signaling in human subcutaneous adipocytes. Finally, the antagonist was established as GIPR selective among 62 human GPCRs being species-specific with high affinity binding to the human and non-human primate (Macaca fascicularis) GIPRs, and low affinity binding to the rat and mouse GIPRs (Kdvalues of 2.0, 2.5, 31.6 and 100 nM, respectively). In conclusion, human GIP(3-30)NH2is a selective and species-specific GIPR antagonist with broad inhibition of signaling and internalization in transfected cells as well as in human adipocytes.",
keywords = "Journal Article",
author = "Gabe, {Maria Buur Nordskov} and Sparre-Ulrich, {Alexander Hovard} and Pedersen, {Mie Fabricius} and Gasbjerg, {L{\ae}rke Smidt} and Asuka Inoue and Hans Br{\"a}uner-Osborne and Bolette Hartmann and Rosenkilde, {Mette Marie}",
note = "Copyright {\circledC} 2018 Elsevier Inc. All rights reserved.",
year = "2018",
month = "1",
day = "31",
doi = "10.1016/j.bcp.2018.01.040",
language = "English",
volume = "150",
pages = "97--107",
journal = "Biochemical Pharmacology",
issn = "0006-2952",
publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - Human GIP(3-30)NH inhibits G protein-dependent as well as G protein-independent signaling and is selective for the GIP receptor with high-affinity binding to primate but not rodent GIP receptors

AU - Gabe, Maria Buur Nordskov

AU - Sparre-Ulrich, Alexander Hovard

AU - Pedersen, Mie Fabricius

AU - Gasbjerg, Lærke Smidt

AU - Inoue, Asuka

AU - Bräuner-Osborne, Hans

AU - Hartmann, Bolette

AU - Rosenkilde, Mette Marie

N1 - Copyright © 2018 Elsevier Inc. All rights reserved.

PY - 2018/1/31

Y1 - 2018/1/31

N2 - GIP(3-30)NH2is a high affinity antagonist of the GIP receptor (GIPR) in humans inhibiting insulin secretion via G protein-dependent pathways. However, its ability to inhibit G protein-independent signaling is unknown. Here we determine its action on arrestin-recruitment and receptor internalization in recombinant cells. As GIP is adipogenic, we evaluate the inhibitory actions of GIP(3-30)NH2in human adipocytes. Finally, we determine the receptor selectivity of GIP(3-30)NH2among other human and animal GPCRs. cAMP accumulation and β-arrestin 1 and 2 recruitment were studied in transiently transfected HEK293 cells and real-time internalization in transiently transfected HEK293A and in HEK293A β-arrestin 1 and 2 knockout cells. Furthermore, human subcutaneous adipocytes were assessed for cAMP accumulation following ligand stimulation. Competition binding was examined in transiently transfected COS-7 cells using human125I-GIP(3-30)NH2. The selectivity of human GIP(3-30)NH2was examined by testing for agonistic and antagonistic properties on 62 human GPCRs. Human GIP(3-30)NH2inhibited GIP(1-42)-induced cAMP and β-arrestin 1 and 2 recruitment on the human GIPR and Schild plot analysis showed competitive antagonism with a pA2and Hill slope of 16.8 nM and 1.11 ± 0.02 in cAMP, 10.6 nM and 1.15 ± 0.05 in β-arrestin 1 recruitment, and 10.2 nM and 1.06 ± 0.05 in β-arrestin 2 recruitment. Efficient internalization of the GIPR was dependent on the presence of either β-arrestin 1 or 2. Moreover, GIP(3-30)NH2inhibited GIP(1-42)-induced internalization in a concentration-dependent manner and notably also inhibited GIP-mediated signaling in human subcutaneous adipocytes. Finally, the antagonist was established as GIPR selective among 62 human GPCRs being species-specific with high affinity binding to the human and non-human primate (Macaca fascicularis) GIPRs, and low affinity binding to the rat and mouse GIPRs (Kdvalues of 2.0, 2.5, 31.6 and 100 nM, respectively). In conclusion, human GIP(3-30)NH2is a selective and species-specific GIPR antagonist with broad inhibition of signaling and internalization in transfected cells as well as in human adipocytes.

AB - GIP(3-30)NH2is a high affinity antagonist of the GIP receptor (GIPR) in humans inhibiting insulin secretion via G protein-dependent pathways. However, its ability to inhibit G protein-independent signaling is unknown. Here we determine its action on arrestin-recruitment and receptor internalization in recombinant cells. As GIP is adipogenic, we evaluate the inhibitory actions of GIP(3-30)NH2in human adipocytes. Finally, we determine the receptor selectivity of GIP(3-30)NH2among other human and animal GPCRs. cAMP accumulation and β-arrestin 1 and 2 recruitment were studied in transiently transfected HEK293 cells and real-time internalization in transiently transfected HEK293A and in HEK293A β-arrestin 1 and 2 knockout cells. Furthermore, human subcutaneous adipocytes were assessed for cAMP accumulation following ligand stimulation. Competition binding was examined in transiently transfected COS-7 cells using human125I-GIP(3-30)NH2. The selectivity of human GIP(3-30)NH2was examined by testing for agonistic and antagonistic properties on 62 human GPCRs. Human GIP(3-30)NH2inhibited GIP(1-42)-induced cAMP and β-arrestin 1 and 2 recruitment on the human GIPR and Schild plot analysis showed competitive antagonism with a pA2and Hill slope of 16.8 nM and 1.11 ± 0.02 in cAMP, 10.6 nM and 1.15 ± 0.05 in β-arrestin 1 recruitment, and 10.2 nM and 1.06 ± 0.05 in β-arrestin 2 recruitment. Efficient internalization of the GIPR was dependent on the presence of either β-arrestin 1 or 2. Moreover, GIP(3-30)NH2inhibited GIP(1-42)-induced internalization in a concentration-dependent manner and notably also inhibited GIP-mediated signaling in human subcutaneous adipocytes. Finally, the antagonist was established as GIPR selective among 62 human GPCRs being species-specific with high affinity binding to the human and non-human primate (Macaca fascicularis) GIPRs, and low affinity binding to the rat and mouse GIPRs (Kdvalues of 2.0, 2.5, 31.6 and 100 nM, respectively). In conclusion, human GIP(3-30)NH2is a selective and species-specific GIPR antagonist with broad inhibition of signaling and internalization in transfected cells as well as in human adipocytes.

KW - Journal Article

U2 - 10.1016/j.bcp.2018.01.040

DO - 10.1016/j.bcp.2018.01.040

M3 - Journal article

C2 - 29378179

VL - 150

SP - 97

EP - 107

JO - Biochemical Pharmacology

JF - Biochemical Pharmacology

SN - 0006-2952

ER -

ID: 189765565