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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


Fluorescence-labeled neuropeptide Y (NPY) has been used in flow cytometric binding assays for the determination of affinity constants of NPY Y1, Y2, and Y5 receptor ligands. Because the binding of fluorescent NPY is insufficient for competition studies at the human Y4 receptor (hY4R), we replaced Glu-4 in hPP with Lys for the derivatization with cyanine-5. Because cy5-[K(4)]hPP has high affinity ...


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