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Molecular Cloning and Characterization of the Rat P2Y 4 Receptor
Author(s) -
Webb Tania E.,
Henderson Duncan J.,
Roberts Jonathan A.,
Barnard Eric A.
Publication year - 1998
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.1998.71041348.x
Subject(s) - receptor , in situ hybridization , biology , messenger rna , microbiology and biotechnology , agonist , open reading frame , molecular cloning , cloning (programming) , stimulation , p2y receptor , endocrinology , biochemistry , gene , complementary dna , peptide sequence , computer science , programming language
Degenerate PCR was used to amplify DNAs encoding members of the P2Y receptor family from rat brain RNA. A full‐length sequence obtained for one novel clone (R5) contained an intronless open reading frame that encoded a polypeptide of 361 amino acids, sharing 84% sequence identity with the human P2Y 4 receptor. When R5 was stably expressed in Jurkat cells, calcium fluxes resulting from stimulation of the receptor showed that UDP, ADP, 2‐methylthio‐ATP, and diadenosine tetraphosphate were inactive, whereas UTP and ATP were both full agonists with similar potency. At the human receptor, ATP has significantly lower potency than UTP. The R5 transcript was not detected in brain by northern hybridization. Therefore, its tissue distribution was assessed by PCR, and the mRNA was found to be widely distributed at a low abundance, being present in brain, spinal cord, and a variety of peripheral organs. Localization of the receptor transcript in adult rat brain sections by in situ hybridization indicated that it is expressed at highest levels in the pineal gland and ventricular system. It is presumed that R5 is a species orthologue of the human P2Y 4 receptor but with this significant difference in agonist pharmacology.