Identification of vasoactive intestinal polypeptide (VIP) binding protein in bovine pineal gland

Abstract Vasoactive intestinal polypeptide (VIP) containing nerves are present in close proximity to epithelial, endocrine, and vascular smooth muscle cells. The pineal gland, known also as a “neuroendocrine transducer organ” contains a high content of VIP which prompted us to characterize the binding sites for VIP in this organ. [Tyr 10 − 125 I]VIP was bound selectively and specifically to pineal membrane preparations in a time-dependent fashion. Scatchard analysis demonstrated a single class of high affinity binding sites with a dissociation constant ( K d ) value of 5.7 ± 0.52 nmol/1 and a receptor density ( B max ) value of 440 ± 35 fmol/mg protein. A Hill Plot with a slope of 1.013 indicated the absence of cooperativity. Covalent crosslinking with [Tyr 10 − 125 I]VIP followed by SDS electrophoresis and autoradiography, revealed that the VIP binding protein exhibited a molecular weight of 51.8 ± 0.5 kDa. The precise function of pineal VIP binding protein remains to be delineated.