Schneider, Erich and Keller, Max and Brennauer, Albert and Hoefelschweiger, Bianca K. and Gross, Dietmar and Wolfbeis, Otto S. and Bernhardt, Günther and Buschauer, Armin (2007) Synthesis and Characterization of the First Fluorescent Nonpeptide NPY Y1 Receptor Antagonist. ChemBioChem 8 (16), pp. 1981-1988.
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Cyanine-5-labelled neuropeptide Y (NPY) was demonstrated to be an ideal universal fluorescent ligand for the combined investigation of NPY Y1, Y2 and Y5 receptors. With respect to improved stability, detection of receptor subtypes in cells and tissues, and prevention of receptor internalization, small nonpeptidic fluorescent antagonists should be superior. Here we present a set of four fluorescent nonpeptide NPY Y1 receptor (Y1R) antagonists. The highest affinity was obtained by labelling an NG-(6-aminohexanoyl)argininamide derived from the Y1R antagonist BIBP 3226, with Py-1, a small pyrylium dye. The fluorescent pyridinium-type Y1R antagonist, compound 4 had Ki values of 29 nM and 2.7 nM, which were determined by radioligand binding and flow cytometry under equilibrium conditions, respectively; 4 had a Kb value of 0.6 nM (Ca2+ assay). The large Stoke's shift (541 vs. 615 nm) in buffer (PBS, pH 7.4) in the presence of 1 % BSA and the red emission (quantum yield 56 %) are advantageous with respect to the signal-to-noise ratio. The new probe was successfully used in fluorescence-based binding experiments evaluated by flow cytometry and confocal microscopy; this demonstrates the potential of pyrylium dyes for the preparation of fluorescent ligands that are applicable for the study of G protein-coupled receptors on living cells.
|Institutions:|| Chemistry and Pharmacy > Institute of Pharmacy > Pharmaceutical/Medicinal Chemistry II (Prof. Buschauer) |
Chemistry and Pharmacy > Institut für Analytische Chemie, Chemo- und Biosensorik > Chemo- und Biosensorik (Prof. Otto S. Wolfbeis)
|Projects:||GRK 760, Graduiertenkolleg Medizinische Chemie|
|Subjects:||500 Science > 570 Life sciences|
600 Technology > 610 Medical sciences Medicine
|Refereed:||Yes, this version has been refereed|
|Created at the University of Regensburg:||Yes|
|Owner:||Prof. Armin Buschauer|
|Deposited On:||21 Feb 2008 07:30|
|Last Modified:||30 Jul 2011 08:27|