Ziemek, Ralf and Schneider, Erich and Kraus, Anja and Cabrele, Chiara and Beck-Sickinger, Annette G. and Bernhardt, Guenther and Buschauer, Armin (2007) Determination of affinity and activity of ligands at the human neuropeptide y Y-4 receptor by flow cytometry and aequorin luminescence. JOURNAL OF RECEPTORS AND SIGNAL TRANSDUCTION, 27 (4). pp. 217-233. ISSN 1079-9893,
Full text not available from this repository. (Request a copy)Abstract
Fluorescence-labeled neuropeptide Y (NPY) has been used in flow cytometric binding assays for the determination of affinity constants of NPY Y-1, Y-2, and Y-5 receptor ligands. Because the binding of fluorescent NPY is insufficient for competition studies at the human Y-4 receptor (hY(4)R), we replaced Glu-4 in hPP with Lys for the derivatization with cyanine-5. Because cy5-[K-4] hPP has high affinity (K-d 5.6 nM) to the hY(4)R, it was used as a probe in a flow cytometric binding assay. Specific binding of cy5-[K-4] hPP to hY(4)R was visualized by confocal microscopy. The hY(4)R, the chimeric G protein G(qi5) and mitochondrially targeted apoaequorin were stably coexpressed in CHO cells. Aequorin luminescence was quantified in a microplate reader and by a CCD camera. By application of these methods 3-cyclohexyl-N-[(3-1H-imidazol-4-ylpropylamino)(imino) methyl] propanamide (UR-AK49) was discovered as the first nonpeptidic Y4R antagonist (pK(i) 4.17), a lead to be optimized in terms of potency and selectivity.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | PANCREATIC-POLYPEPTIDE RECEPTORS; PROTEIN-COUPLED RECEPTORS; FUNCTIONAL CALCIUM ASSAY; FOOD-INTAKE; NEUROKININ RECEPTORS; NPY RECEPTORS; PEPTIDE YY; CELL-LINES; AGONIST; RAT; NPYY4 receptor; flow cytometry; chimeric g protein; Aequorin; nonpeptide NPYY4 receptor antagonist |
| Subjects: | 600 Technology > 615 Pharmacy |
| Divisions: | Chemistry and Pharmacy > Institute of Pharmacy > Alumni or Retired Professors > Pharmaceutical/Medicinal Chemistry II (Prof. Buschauer) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 12 Jan 2021 10:58 |
| Last Modified: | 12 Jan 2021 10:58 |
| URI: | https://pred.uni-regensburg.de/id/eprint/33463 |
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