N- and C-terminally truncated forms of glucose-dependent insulinotropic polypeptide are high-affinity competitive antagonists of the human GIP receptor

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N- and C-terminally truncated forms of glucose-dependent insulinotropic polypeptide are high-affinity competitive antagonists of the human GIP receptor. / Hansen, L S; Sparre-Ulrich, A H; Christensen, M.; Knop, F K; Hartmann, B; Holst, J J; Rosenkilde, M M.

In: British Journal of Pharmacology, Vol. 173, No. 5, 03.2016, p. 826-838.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Hansen, LS, Sparre-Ulrich, AH, Christensen, M, Knop, FK, Hartmann, B, Holst, JJ & Rosenkilde, MM 2016, 'N- and C-terminally truncated forms of glucose-dependent insulinotropic polypeptide are high-affinity competitive antagonists of the human GIP receptor', British Journal of Pharmacology, vol. 173, no. 5, pp. 826-838. https://doi.org/10.1111/bph.13384

APA

Hansen, L. S., Sparre-Ulrich, A. H., Christensen, M., Knop, F. K., Hartmann, B., Holst, J. J., & Rosenkilde, M. M. (2016). N- and C-terminally truncated forms of glucose-dependent insulinotropic polypeptide are high-affinity competitive antagonists of the human GIP receptor. British Journal of Pharmacology, 173(5), 826-838. https://doi.org/10.1111/bph.13384

Vancouver

Hansen LS, Sparre-Ulrich AH, Christensen M, Knop FK, Hartmann B, Holst JJ et al. N- and C-terminally truncated forms of glucose-dependent insulinotropic polypeptide are high-affinity competitive antagonists of the human GIP receptor. British Journal of Pharmacology. 2016 Mar;173(5):826-838. https://doi.org/10.1111/bph.13384

Author

Hansen, L S ; Sparre-Ulrich, A H ; Christensen, M. ; Knop, F K ; Hartmann, B ; Holst, J J ; Rosenkilde, M M. / N- and C-terminally truncated forms of glucose-dependent insulinotropic polypeptide are high-affinity competitive antagonists of the human GIP receptor. In: British Journal of Pharmacology. 2016 ; Vol. 173, No. 5. pp. 826-838.

Bibtex

@article{5043b16ed0954774b9bdb42066678e24,
title = "N- and C-terminally truncated forms of glucose-dependent insulinotropic polypeptide are high-affinity competitive antagonists of the human GIP receptor",
abstract = "BACKGROUND AND PURPOSE: Glucose-dependent insulinotropic polypeptide (GIP) impacts lipid, bone, and glucose homeostasis. The GIP receptor belongs to G protein-coupled receptor family B1 and signals through GαS. High affinity ligands for in vivo use are needed to elucidate GIP's physiological functions and pharmacological potential. GIP(1-30)NH2 is a naturally occurring truncation of GIP(1-42). Here we characterize eight N-terminal trrncations of human GIP(1-30)NH2 : GIP(2- to 9-30)NH2 .EXPERIMENTAL APPROACH: COS-7 cells were transiently transfected with the human GIP receptor and assessed for cAMP accumulation upon ligand stimulation or competition binding with (125) I-GIP(1-42), (125) I-GIP(1-30)NH2 , (125) I-GIP(2-30)NH2 , or (125) I-GIP(3-30)NH2 as radioligands.KEY RESULTS: GIP(1-30)NH2 displaced (125) I-GIP(1-42) equally to GIP(1-42) (Ki 0.75 nM), whereas the eight variants displayed lower affinities (Ki 2.3-347 nM) with highest affinities of GIP(3-30)NH2 and (5-30)NH2 . Agonism was only observed for GIP(1-30)NH2 with an Emax on 100% of GIP(1-42) and GIP(2-30)NH2 (Emax 20%). GIP(2- to 9-30)NH2 displayed antagonism (IC50 12-450 nM) and right-shifts of the GIP(1-42)-response curve. Schild plot analyses identified GIP(3-30)NH2 and GIP(5-30)NH2 as competitive antagonists (Ki 15 nM). Importantly, GIP(3-30) antagonized with a 26-fold higher potency than GIP(3-42). Binding studies with agonist ((125) I-GIP(1-30)NH2 ), partial agonist ((125) I-GIP(2-30)NH2 ) and competitive antagonist ((125) I-GIP(3-30)NH2 ) revealed distinct receptor conformations for these three ligand classes.CONCLUSIONS AND IMPLICATIONS: The N-terminus is crucial for GIP agonist functionality. Removal of the C-terminus of the naturally occurring DPP4-product GIP(3-42) creates another naturally occurring, but superior antagonist GIP(3-30)NH2 , that together with GIP(5-30)NH2 were high-affinity competitive antagonist and thus may be suitable tool compounds for basic GIP research and future pharmacological interventions.",
author = "Hansen, {L S} and Sparre-Ulrich, {A H} and M. Christensen and Knop, {F K} and B Hartmann and Holst, {J J} and Rosenkilde, {M M}",
note = "This article is protected by copyright. All rights reserved.",
year = "2016",
month = mar,
doi = "10.1111/bph.13384",
language = "English",
volume = "173",
pages = "826--838",
journal = "British Journal of Pharmacology",
issn = "0007-1188",
publisher = "Wiley",
number = "5",

}

RIS

TY - JOUR

T1 - N- and C-terminally truncated forms of glucose-dependent insulinotropic polypeptide are high-affinity competitive antagonists of the human GIP receptor

AU - Hansen, L S

AU - Sparre-Ulrich, A H

AU - Christensen, M.

AU - Knop, F K

AU - Hartmann, B

AU - Holst, J J

AU - Rosenkilde, M M

N1 - This article is protected by copyright. All rights reserved.

PY - 2016/3

Y1 - 2016/3

N2 - BACKGROUND AND PURPOSE: Glucose-dependent insulinotropic polypeptide (GIP) impacts lipid, bone, and glucose homeostasis. The GIP receptor belongs to G protein-coupled receptor family B1 and signals through GαS. High affinity ligands for in vivo use are needed to elucidate GIP's physiological functions and pharmacological potential. GIP(1-30)NH2 is a naturally occurring truncation of GIP(1-42). Here we characterize eight N-terminal trrncations of human GIP(1-30)NH2 : GIP(2- to 9-30)NH2 .EXPERIMENTAL APPROACH: COS-7 cells were transiently transfected with the human GIP receptor and assessed for cAMP accumulation upon ligand stimulation or competition binding with (125) I-GIP(1-42), (125) I-GIP(1-30)NH2 , (125) I-GIP(2-30)NH2 , or (125) I-GIP(3-30)NH2 as radioligands.KEY RESULTS: GIP(1-30)NH2 displaced (125) I-GIP(1-42) equally to GIP(1-42) (Ki 0.75 nM), whereas the eight variants displayed lower affinities (Ki 2.3-347 nM) with highest affinities of GIP(3-30)NH2 and (5-30)NH2 . Agonism was only observed for GIP(1-30)NH2 with an Emax on 100% of GIP(1-42) and GIP(2-30)NH2 (Emax 20%). GIP(2- to 9-30)NH2 displayed antagonism (IC50 12-450 nM) and right-shifts of the GIP(1-42)-response curve. Schild plot analyses identified GIP(3-30)NH2 and GIP(5-30)NH2 as competitive antagonists (Ki 15 nM). Importantly, GIP(3-30) antagonized with a 26-fold higher potency than GIP(3-42). Binding studies with agonist ((125) I-GIP(1-30)NH2 ), partial agonist ((125) I-GIP(2-30)NH2 ) and competitive antagonist ((125) I-GIP(3-30)NH2 ) revealed distinct receptor conformations for these three ligand classes.CONCLUSIONS AND IMPLICATIONS: The N-terminus is crucial for GIP agonist functionality. Removal of the C-terminus of the naturally occurring DPP4-product GIP(3-42) creates another naturally occurring, but superior antagonist GIP(3-30)NH2 , that together with GIP(5-30)NH2 were high-affinity competitive antagonist and thus may be suitable tool compounds for basic GIP research and future pharmacological interventions.

AB - BACKGROUND AND PURPOSE: Glucose-dependent insulinotropic polypeptide (GIP) impacts lipid, bone, and glucose homeostasis. The GIP receptor belongs to G protein-coupled receptor family B1 and signals through GαS. High affinity ligands for in vivo use are needed to elucidate GIP's physiological functions and pharmacological potential. GIP(1-30)NH2 is a naturally occurring truncation of GIP(1-42). Here we characterize eight N-terminal trrncations of human GIP(1-30)NH2 : GIP(2- to 9-30)NH2 .EXPERIMENTAL APPROACH: COS-7 cells were transiently transfected with the human GIP receptor and assessed for cAMP accumulation upon ligand stimulation or competition binding with (125) I-GIP(1-42), (125) I-GIP(1-30)NH2 , (125) I-GIP(2-30)NH2 , or (125) I-GIP(3-30)NH2 as radioligands.KEY RESULTS: GIP(1-30)NH2 displaced (125) I-GIP(1-42) equally to GIP(1-42) (Ki 0.75 nM), whereas the eight variants displayed lower affinities (Ki 2.3-347 nM) with highest affinities of GIP(3-30)NH2 and (5-30)NH2 . Agonism was only observed for GIP(1-30)NH2 with an Emax on 100% of GIP(1-42) and GIP(2-30)NH2 (Emax 20%). GIP(2- to 9-30)NH2 displayed antagonism (IC50 12-450 nM) and right-shifts of the GIP(1-42)-response curve. Schild plot analyses identified GIP(3-30)NH2 and GIP(5-30)NH2 as competitive antagonists (Ki 15 nM). Importantly, GIP(3-30) antagonized with a 26-fold higher potency than GIP(3-42). Binding studies with agonist ((125) I-GIP(1-30)NH2 ), partial agonist ((125) I-GIP(2-30)NH2 ) and competitive antagonist ((125) I-GIP(3-30)NH2 ) revealed distinct receptor conformations for these three ligand classes.CONCLUSIONS AND IMPLICATIONS: The N-terminus is crucial for GIP agonist functionality. Removal of the C-terminus of the naturally occurring DPP4-product GIP(3-42) creates another naturally occurring, but superior antagonist GIP(3-30)NH2 , that together with GIP(5-30)NH2 were high-affinity competitive antagonist and thus may be suitable tool compounds for basic GIP research and future pharmacological interventions.

U2 - 10.1111/bph.13384

DO - 10.1111/bph.13384

M3 - Journal article

C2 - 26572091

VL - 173

SP - 826

EP - 838

JO - British Journal of Pharmacology

JF - British Journal of Pharmacology

SN - 0007-1188

IS - 5

ER -

ID: 150704907