High-affinity binding of [ 3H]neuropeptide Y to a polypeptide from the venom of Conus anemone

• Published in: European journal of pharmacology Vol. 315; no. 3; pp. 355 - 362
• Main Authors: Czerwiec, Eva, De Backer, Jean-Paul, Vauquelin, Georges, Vanderheyden, Patrick M.L.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 21.11.1996; Elsevier
• Subjects: Animal poisons toxicology. Antivenoms; Animals; Binding protein; Binding, Competitive; Biological and medical sciences; Cattle; Cell receptors; Cell structures and functions; Cerebral Cortex - drug effects; Conus anemone; Dose-Response Relationship, Drug; Fundamental and applied biological sciences. Psychology; Humans; Medical sciences; Molecular and cellular biology; Neuropeptide receptors; Neuropeptide Y (NPY); Neuropeptide Y - metabolism; Neuropeptide Y-(18-36); Pancreatic polypeptide; Peptides - metabolism; Rats; Toxicology; Venoms - metabolism; Venoms - pharmacology; Binding protein; Neuropeptide Y (NPY); Pancreatic polypeptide; Neuropeptide Y-(18-36); Conus anemone; Neuropeptide Y; Toxicity; Animal origin; Molecular interaction; Identification; Gastropoda; Conidae; In vitro; Toxin; Biological fixation; Structure activity relation; Venom; Peptidergic receptor; Affinity; Invertebrata; Mollusca; Localization; Mechanism of action
• ISSN: 0014-2999; 1879-0712
• DOI: 10.1016/S0014-2999(96)00647-4

Venom preparation from Conus anemone contains a component that binds radiolabeled neuropeptide Y ([ 3H]neuropeptide Y) with high affinity ( K D = 2.9 nM ± 0.2 nM, B max = 15.2 ± 0.5 pmol/mg protein). Binding of [ 3H]neuropeptide Y to the venom component is displaced with nanomolar affinity of unlabeled human and porcine neuropeptide Y, porcine [Leu 31-Pro 34]neuropeptide Y, peptide YY, avian and bovine pancreatic polypeptide, and the (18–36) and (25–36) C-terminal fragments from neuropeptide Y. No displacement is found with the (1–24) N-terminal neuropeptide Y fragment, human secretin, porcine dynorphin A and Boc-DAKLI (Bolton Hunter coupled dynorphin A analog kappa ligand) nor with the non-peptide neuropeptide Y receptor antagonist BIBP3266. Gel filtration chromatography and denaturing (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) show that the [ 3H]neuropeptide Y-binding component is very likely a single-chain polypeptide with a molecular mass of 18.5 kDa.