Novel cell line selectively expressing neuropeptide Y-Y2 receptors.
Neuropeptide Y (NPY) recognition by the human neuroblastoma cell lines SiMa, Kelly, SH-SY5Y, CHP-234, and MHH-NB-11 was analyzed in radioactive binding assays using tritiated NPY. For the cell lines CHP-234 and MHH-NB-11 binding of [3H]propionyl-NPY was observed with Kd-values of 0.64 +/- 0.07 nM and 0.53 +/- 0.12 nM, respectively, determined by saturation analysis with non-linear regression. The receptor subtype was determined by competition analysis using the subtype selective NPY analogues [Leu31, Pro34]-NPY (NPY-Y1, NPY-Y5), [Ahx(5-24)]-NPY (NPY-Y2), [Ala31, Aib32]-NPY (NPY-Y5), NPY [3-36] (NPY-Y2, NPY-Y5), and NPY [13-36] (NPY-Y2). Both cell lines, CHP-234 and MHH-NB-11, the latter one being characterized for NPY receptors for the first time, showed exclusive expression of NPY-Y2 receptors. In both cell lines binding of NPY induced signal transduction, which was monitored as reduction of forskolin-induced cAMP production in an ELISA.
Höfliger
Martin M
MM
Institute of Biochemistry, University of Leipzig, Leipzig, Germany.
Castejón
Gloria López
GL
Kiess
Wieland
W
Beck Sickinger
Annette G
AG
D000595:Amino Acid Sequence; D001667:Binding, Competitive; D045744:Cell Line, Tumor; D000242:Cyclic AMP / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007700:Kinetics; D016014:Linear Models; D008969:Molecular Sequence Data; D010452:Peptide Biosynthesis; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D017476:Receptors, Neuropeptide Y / Q000096:biosynthesis*; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction
D004791:Enzyme Inhibitors; D010455:Peptides; D017476:Receptors, Neuropeptide Y; C095889:neuropeptide Y2 receptor; D000242:Cyclic AMP