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Determination of affinity and activity of ligands at the human neuropeptide Y Y4 receptor by flow cytometry and aequorin luminescence.

Ziemek, Ralf ; Schneider, Erich ; Kraus, Anja ; Cabrele, Chiara ; Beck-Sickinger, Annette G. ; Bernhardt, Günther ; Buschauer, Armin



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 ...

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