RRC ID 48103
Author Masu Y, Nakayama K, Tamaki H, Harada Y, Kuno M, Nakanishi S.
Title cDNA cloning of bovine substance-K receptor through oocyte expression system.
Journal Nature
Abstract The neuropeptide receptors which are present in very small quantities in the cell and are embedded tightly in the plasma membrane have not been well characterized. Mammals contain three distinct tachykinin neuropeptides, substance P, substance K and neuromedin K, and it has been suggested that there are multiple tachykinin receptors. By electrophysiological measurement, we have previously shown that Xenopus oocytes injected with brain and stomach mRNAs faithfully express mammalian substance-P and substance-K receptors, respectively. Here we report the isolation of the cDNA clone for bovine substance-K receptor (SKR) by extending this method to develop a new cloning strategy. We constructed a stomach cDNA library with a cloning vector that allowed in vitro synthesis of mRNAs and then identified a particular cDNA clone by testing for receptor expression following injection of the mRNAs synthesized in vitro into the oocyte system. Because oocytes injected with exogenous mRNAs can express numerous receptors and channels, our new strategy will be applicable in the general molecular cloning of these proteins. The result provides the first indication that the neuropeptide receptor has sequence similarity with rhodopsin-type receptors (the G-protein-coupled receptor family) and thus possesses multiple membrane-spanning domains.
Volume 329(6142)
Pages 836-8
Published 1987-10-29
DOI 10.1038/329836a0
PMID 2823146
MeSH Amino Acid Sequence Animals Base Sequence Cattle Cloning, Molecular* DNA / metabolism* Female Molecular Sequence Data Oocytes / metabolism Promoter Regions, Genetic Receptors, Neurokinin-2 Receptors, Neurotransmitter / genetics* Sequence Homology, Nucleic Acid Transcription, Genetic*
IF 42.779
Times Cited 700
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
DNA material pbSKR 56 S (RDB13188) pbSKR 56 T (RDB13189).