Cloning and functional expression of cDNAs encoding human and rat pancreatic polypeptide receptors.
PCR was used to isolate nucleotide sequences that may encode novel members of the neuropeptide Y receptor family. By use of a PCR product as a hybridization probe, a full-length human cDNA was isolated that encodes a 375-aa protein with a predicted membrane topology identifying it as a member of the G-protein-coupled receptor superfamily. After stable transfection of the cDNA into human embryonic kidney 293 cells, the receptor exhibited high affinity (Kd = 2.8 nM) for 125I-labeled human pancreatic polypeptide (PP). Competition binding studies in whole cells indicated the following rank order of potency: human PP = bovine PP > or = human [Pro34]peptide YY > rat PP > human peptide YY = human neuropeptide Y. Northern blot analysis revealed that human PP receptor mRNA is most abundantly expressed in skeletal muscle and, to a lesser extent, in lung and brain tissue. A rat cDNA clone encoding a high-affinity PP receptor that is 74% identical to the human PP receptor at the amino acid level was also isolated. These receptor clones will be useful in elucidating the functional role of PP and designing selective PP receptor agonists and antagonists.
Yan
H
H
Department of Neurobiology, Amgen Center, Thousand Oaks, CA 91320, USA.
Yang
J
J
Marasco
J
J
Yamaguchi
K
K
Brenner
S
S
Collins
F
F
Karbon
W
W
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D002417:Cattle; D002460:Cell Line; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D009478:Neuropeptide Y / Q000378:metabolism; D010191:Pancreatic Polypeptide / Q000378:metabolism*; D019894:Peptide YY; D010455:Peptides / Q000378:metabolism; D016133:Polymerase Chain Reaction; D051381:Rats; D011964:Receptors, Gastrointestinal Hormone / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014162:Transfection
D017931:DNA Primers; D018076:DNA, Complementary; D009478:Neuropeptide Y; D010455:Peptides; D011964:Receptors, Gastrointestinal Hormone; D011994:Recombinant Proteins; C029438:receptors, pancreatic polypeptide-specific; D019894:Peptide YY; D010191:Pancreatic Polypeptide