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
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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