A novel truncated glucagon-like peptide 2 (GLP-2) as a tool for analyzing GLP-2 receptor agonists

Yamazaki, Kazuto; Kagaya, Toshitaka; Watanabe, Misako; Terauchi, Hiroki; Iida, Daisuke; Fukumoto, Hironori; Suzuki, Shinichi; Arai, Tohru; Aoki, Mika; Takase, Kazuma; Seiki, Takashi; Tsukahara, Kappei; Nagakawa, Junichi

doi:10.2220/biomedres.34.129

Cited by 11 publications

(15 citation statements)

References 24 publications

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“…The binding interface with the extracellular domain (ECD) of the receptor was predicted to be between Leucine17 and Lysine30, while the N-terminus part of GLP-2 from Histiine1 to Aspargine16 lacked contact with the extracellular domains of the GLP-2R. The central roles of the N-and C-terminus part of GLP-2 in respectively, receptor activation and receptor binding were supported by Yamazaki et. al.…”

Section: Discussionmentioning

confidence: 90%

“…al. in 2013(Yamazaki et al 2013, showing a decreased intrinsic placental alkaline phosphatase (PALP) activity (driven by cAMP) for GLP-2(3-33), (6-33) and (11-to 13-33). Most recently, Wisniewski et.…”

Section: Discussionmentioning

confidence: 99%

“…These changes can have profound structural and functional consequences (Chao et al 1997;Hoshi et al 2001;Gu et al 2015;Sugamura et al 2011). To protect for oxidative damage of the Met in GLP-2 during the oxidative iodination, and since Met is dispensable for GLP-2 function (Drucker et al 2013;Venneti et al 2011;Wisniewski et al 2016;Yamazaki et al 2013), we replaced Met10 with a Tyr residue. Thereby we created a target site for oxidative iodination using [ 125 I] in the full agonist (GLP-2(1-33)) and in the antagonist and partial agonist (GLP-2(3-33)).…”

Section: Discussionmentioning

confidence: 99%

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Development of high-affinity agonist- and antagonist radioligands for the GLP-2 receptor - powerful tools for the study of GLP-2 pharmacology

Gadgaard

Velden

Schiellerup

et al. 2020

Preprint

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Background: Glucagon-like peptide-2 (GLP-2) is a 33 amino acid pro-glucagon-derived hormone produced in the intestinal enteroendocrine L-cells with trophic actions on both the gut and bones. GLP-2(1-33) is cleaved by the ubiquitous protease dipeptidyl peptidase-4 (DPP-4), resulting in with competitive antagonistic properties on the GLP-2 receptor (GLP-2R). Here we present two new hGLP-2 radioligands with different pharmacodynamic profiles. Experimental Approach: The methionine in position 10 of GLP-2(1-33) was substituted with tyrosine to enable oxidative iodination with incorporation of the iodine isotope [125I]. Similar substitution was done in GLP-2(3-33), thereby creating two new radioligands; an agonist [125]-hGLP-2(1-33,M10Y) and an antagonist [125]-hGLP-2(3-33,M10Y). Both were characterized regarding competition binding, binding kinetics and target tissue autoradiography. Key results: High and similar binding affinities for the human GLP-2R were observed for [125I]-hGLP-2(1-33,M10Y) and [125I]-hGLP-2(3-33,M10Y) with KD values of 59.3 nM and 40.6 nM, respectively. The M10Y substitution did not change the functional properties of GLP-2(1-33) or GLP-2(3-33). The antagonist [125I]-hGLP-2(3-33,M10Y) had higher Bmax and faster on-rate for the hGLP-2R compared to the agonist [125I]-hGLP-2(1-33,M10Y). Using autoradiography in mice strong labeling was observed in subepithelial myofibroblasts (SEMF) and pancreas islet cells. Both radioligands were selective for the GLP-2R, except for a low affinity binding to the GLP-1R (IC50 of 130 and 330 nM, respectively) Conclusion and implications: We successfully developed two new high affinity radioligands for GLP-2R studies and identified SEMF and pancreatic islets as target for GLP-2. It is uncertain whether binding in the pancreas islets results from GLP-2R or GLP-1R binding.

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Section: Discussionmentioning

confidence: 90%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Development of high-affinity agonist- and antagonist radioligands for the GLP-2 receptor - powerful tools for the study of GLP-2 pharmacology

Gadgaard

Velden

Schiellerup

et al. 2020

Preprint

View full text Add to dashboard Cite

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“…These changes can have profound structural and functional consequences (Gu et al, 2015;Hoshi & Heinemann, 2001). To protect for oxidative damage of the Met in GLP-2 during the oxidative iodination, and since Met is dispensable for GLP-2 function (Drucker et al, 1996;Venneti & Hewage, 2011;Wi sniewski et al, 2016;Yamazaki et al, 2013), we replaced Met10 with a Tyr residue. Thereby, we created a target site for oxidative iodination using the binding and activation of both the GLP-1 receptor and the glucagon receptor by oxyntomodulin (Holst et al, 2018;Jorgensen et al, 2007) and the activation of the GLP-1 receptor by glucagon (Svendsen et al, 2018).…”

Section: Selectively Binding Profile Of the Two Radioligands For Rela...mentioning

confidence: 99%

Novel agonist and antagonist radioligands for the GLP‐2 receptor. Useful tools for studies of basic GLP‐2 receptor pharmacology

Gadgaard

Velden

Schiellerup

et al. 2022

British J Pharmacology

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Background Glucagon‐like peptide‐2 (GLP‐2) is a pro‐glucagon‐derived hormone secreted from intestinal enteroendocrine L cells with actions on gut and bones. GLP‐2(1–33) is cleaved by DPP‐4, forming GLP‐2(3–33), having low intrinsic activity and competitive antagonism properties at GLP‐2 receptors. We created radioligands based on these two molecules. Experimental approach The methionine in position 10 of GLP‐2(1–33) and GLP‐2(3–33) was substituted with tyrosine (M10Y) enabling oxidative iodination, creating [125I]‐hGLP‐2(1–33,M10Y) and [125I]‐hGLP‐2(3–33,M10Y). Both were characterized by competition binding, on‐and‐off‐rate determination and receptor activation. Receptor expression was determined by target‐tissue autoradiography and immunohistochemistry. Key results Both M10Y‐substituted peptides induced cAMP production via the GLP‐2 receptor comparable to the wildtype peptides. GLP‐2(3–33,M10Y) maintained the antagonistic properties of GLP‐2(3–33). However, hGLP‐2(1–33,M10Y) had lower arrestin recruitment than hGLP‐2(1–33). High affinities for the hGLP‐2 receptor were observed using [125I]‐hGLP‐2(1–33,M10Y) and [125I]‐hGLP‐2(3–33,M10Y) with KD values of 59.3 and 40.6 nM. The latter (with antagonistic properties) had higher Bmax and faster on and off rates compared to the former (full agonist). Both bound the hGLP‐1 receptor with low affinity (Ki of 130 and 330 nM, respectively). Autoradiography in wildtype mice revealed strong labelling of subepithelial myofibroblasts, confirmed by immunohistochemistry using a GLP‐2 receptor specific antibody that in turn was confirmed in GLP‐2 receptor knock‐out mice. Conclusion and implications Two new radioligands with different binding kinetics, one a full agonist and the other a weak partial agonist with antagonistic properties were developed and subepithelial myofibroblasts identified as a major site for GLP‐2 receptor expression.

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“…The mechanisms linking fat accumulation to bone health are unclear. Therefore, we investigated some physiological actions of GIP and GLP-2 in rats by subchronic treatment with a novel GIPR antagonist, GIP(3-30)NH 2 (14, 22), a partial agonist, (Pro3)GIP and two different GLP-2R antagonists [GLP-2(11-33), and GLP-2(3-33)] (31, 46, 47) in an attempt to (1) evaluate their action in the regulation of lipid and bone homeostasis and (2) identify potential mechanisms linking the two together.…”

Section: Introductionmentioning

confidence: 99%

Increased Body Weight and Fat Mass After Subchronic GIP Receptor Antagonist, but Not GLP-2 Receptor Antagonist, Administration in Rats

et al. 2019

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Glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-2 (GLP-2) are hormones secreted from the enteroendocrine cells after a meal. They exert their actions through activation of G protein-coupled receptors (R), the GIPR and GLP-2R, respectively. Both have been reported to influence metabolism. The purpose of the study was to investigate the role of the hormones in the regulation of lipid and bone homeostasis by subchronic treatment with novel GIPR and GLP-2R antagonists. Rats were injected once daily with vehicle, GIPR, or GLP-2R antagonists for 3 weeks. Body weight, food intake, body composition, plasma lipoprotein lipase (LPL), adipokines, triglycerides and the marker of bone resorption carboxy-terminal collagen crosslinks (CTX), were examined. In rats, subchronic treatment with GIPR antagonist, rat GIP (3-30)NH 2 , did not modify food intake and bone resorption, but significantly increased body weight, body fat mass, triglycerides, LPL, and leptin levels compared with vehicle treated rats. Subchronic (Pro3)GIP (a partial GIPR agonist), GLP-2(11-33), and GLP-2(3-33) (GLP-2R antagonists) treatment did not affect any parameter. The present results would be consistent with a role for GIP, but not GLP-2, in the maintenance of lipid homeostasis in rats, while neither GIPR nor GLP-2R antagonism appeared to influence bone resorption in rats.

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A novel truncated glucagon-like peptide 2 (GLP-2) as a tool for analyzing GLP-2 receptor agonists

Cited by 11 publications

References 24 publications

Development of high-affinity agonist- and antagonist radioligands for the GLP-2 receptor - powerful tools for the study of GLP-2 pharmacology

Development of high-affinity agonist- and antagonist radioligands for the GLP-2 receptor - powerful tools for the study of GLP-2 pharmacology

Novel agonist and antagonist radioligands for the GLP‐2 receptor. Useful tools for studies of basic GLP‐2 receptor pharmacology

Increased Body Weight and Fat Mass After Subchronic GIP Receptor Antagonist, but Not GLP-2 Receptor Antagonist, Administration in Rats

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