Coexpression with potassium channel subunits used to clone the Y2 receptor for neuropeptide Y.
Xenopus oocytes were injected with RNAs for the two inward-rectifier potassium channel subunits Kir3.1 (GIRK1) and Kir3.4 (rcKATP or CIR) in addition to RNA from the neuroblastoma cell line KAN-TS. Potassium currents were evoked by neuropeptide Y in oocytes injected with polyadenylated RNA or with cRNA from pools of a neuroblastoma (KAN-TS) cDNA library, and progressive subdivision of responding pools yielded a single cDNA. The encoded protein contains 381 amino acids, has the seven hydrophobic domains characteristic of G protein-coupled receptors, and is 31% identical to the Y1 receptor: potassium currents were induced by neuropeptide Y (EC50=60pm) and Y2-selective analogues. Coexpression with potassium channel subunits will be a generally useful method for the cloning of G protein-coupled receptors.
Rimland
J M
JM
Glaxo Research Laboratories, Verona, Italy.
Seward
E P
EP
Humbert
Y
Y
Ratti
E
E
Trist
D G
DG
North
R A
RA
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D006801:Humans; D008969:Molecular Sequence Data; D009447:Neuroblastoma; D009478:Neuropeptide Y / Q000378:metabolism* / Q000494:pharmacology; D009865:Oocytes / Q000502:physiology / Q000648:ultrastructure; D015221:Potassium Channels / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D012313:RNA / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D017476:Receptors, Neuropeptide Y / Q000235:genetics* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014407:Tumor Cells, Cultured; D014981:Xenopus
D009478:Neuropeptide Y; D015221:Potassium Channels; D012333:RNA, Messenger; D017476:Receptors, Neuropeptide Y; C095889:neuropeptide Y2 receptor; D012313:RNA