Journal of Biological Chemistry · 1995 · 272 citations · 37 references
Life SciencesNeurosciencePharmacologyMolecular NeuroscienceNeuroendocrinologyNeuropharmacologyNeuropeptide PharmacologyNeurotransmissionNeurotransmitter ReceptorsPancreatic HormonesReceptor PharmacologyReceptor PhysiologyCdna Library ConstructionNeuropeptide Receptor PharmacologyNeuropeptide Receptor SubtypesNeuropeptide Y PharmacologyPancreatic Peptide HormonesPeptide Hormone PharmacologyPeptide Hormone ReceptorsReceptors
The pancreatic polypeptide family includes neuropeptide Y (NPY), one of the most abundant neuropeptides in the mammalian nervous system, as well as peptide YY (PYY) and pancreatic polypeptide (PP). This peptide family is involved in numerous physiological processes such as memory, pain, blood pressure, appetite, anxiety, and circadian rhythm. Of the multiple Y-type receptors proposed for PP family members, only the Y1 subtype was cloned previously. We now report the isolation of a human Y2 (hhY2) receptor cDNA by expression cloning from a human hippocampal cDNA library, using a 125I-PYY binding assay. hhY2 cDNA encodes a predicted protein of 381 amino acids with low amino acid identity to the human Y1 receptor (31% overall; 41% transmembrane). 125I-PYY binding to transiently expressed hY2 receptors was saturable (pKd = 10.17) and displaceable by human PP family members in rank order: PYY (pKi = 9.47) approximately NPY (pKi = 9.27) >> PP (pKi < 6) and by peptide analogs: NPY2-36 (pKi = 8.80) approximately NPY13-36 (pKi = 8.55) approximately C2-NPY (pKi = 8.54) > NPY26-36 (pKi = 6.51) approximately [Leu31,Pro34]NPY (pKi = 6.23). Human PYY decreased [cAMP] and increased intracellular [Ca2+] in hY2-transfected 293 cells.
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Molecular cloning: A laboratory manual
Prescott L. Deininger · Analytical Biochemistry · 1990 · 86.2K citations
A simple and very efficient method for generating cDNA libraries
Ueli Gubler, Beth J. Hoffman · Gene · 1983 · 4.6K citations