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Characterization of specific calcitonin gene related peptide receptors present in hamster pancreatic β cells

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Bioscience Reports

Abstract

Calcitonin gene-related peptide (CGRP) shares about 46% and 20% amino acid sequence homology with islet amyloid polypeptide (IAPP) and salmon calcitonin (sCT). We investigated whether these related peptides could cross-react with the specific binding of125I-[His]hCGRP I to the CGRP receptor in hamster insulinoma cell membranes. A rapid dissociation of membrane bound125I-[His]hCGRP I could be induced in the presence of 1 μM chicken CGRP (cCGRP). The specific125I-[His]hCGRP I binding was inhibited by the related peptides and their half-maximal inhibitory concentrations (IC50) were: cCGRP (0.1 nM), rat CGRP I and human CGRP I and II (1.0–2.0 nM), fragment of hCGRP I (8-37) (150 nM), human IAPP (440 nM). The non-amidated form of hIAPP; human diabetes-associated peptide (hDAP) did not inhibit the binding of125I-[His]hCGRP I and sCT was only effective at a high concentration (1 μM). Binding of125I-[His]hCGRP I was dose dependently inhibited by guanosine-5′-O-(3-thiotriphosphate) or (GTPγS) and a 70% reduction of binding was obtained with 0.1 mM GTPγS. The IC50 value of cCGRP (0.1 nM) was increased 100-fold in the presence of 0.1 mM GTPγS. Human CGRP I and cCGRP at 2.5 μM did not stimulate the activity of hamster insulinoma cell membranes adenylate cyclase, while glucagon (1 μM) induced a 2-fold increase. Thus, specific CGRP receptors present in hamster β cells are associated with G protein (s) and IAPP can interact with these receptors. These results and the observation that cCGRP and hCGRP I did not influence adenylate cyclase activity provide further evidence for CGRP receptor subtypes.

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Abbreviations

CGRP:

calcitonin gene-related peptide

IAPP:

islet amyloid polypeptide

IC50 :

half-maximal inhibitory concentration

GTPγS:

guanosine-5′-O-(3-thiotriphosphate)

125I:

[His]hCGRP I, (2[125I]iodohistidyl10) human CGRP I

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Authors and Affiliations

  1. Institut National de la Santé et de la Recherche Médicale U.55, Hôpital Saint-Antoine, 75571, Paris Cedex 12, France

    A. Barakat, G. Rosselin & J. -C. Marie

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  1. A. Barakat
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  2. G. Rosselin
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  3. J. -C. Marie
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Barakat, A., Rosselin, G. & Marie, J.C. Characterization of specific calcitonin gene related peptide receptors present in hamster pancreatic β cells. Biosci Rep 13, 221–231 (1993). https://doi.org/10.1007/BF01123504

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  • DOI: https://doi.org/10.1007/BF01123504

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