Molecular cloning and characterization of the mouse P2Y4 nucleotide receptor.
To isolate the mouse P2Y4 receptor gene, a mouse genomic library was screened with a human P2Y4 probe. An open reading frame encoding a protein of 361 amino acids was isolated. This protein showed 82% and 95% amino acid identity with the human and rat P2Y4 receptors, respectively. By reverse transcription and polymerase chain reaction (RT-PCR), the P2Y4 messenger RNA was detected in mouse liver, intestine, stomach, bladder and lung among the 16 mouse tissues tested. In 1321N1 transfected cells, the mouse P2Y4 receptor was equally activated by UTP and ATP, and was antagonized by pyridoxal-phosphate-6-azophenyl-2',4'-disulphonic acid (PPADS) and Reactive Blue 2, and not by suramin. Moreover, when expressed in 1321N1 cells, the rat P2Y4 is also antagonized by PPADS. Thus, when compared in the same expression system, the mouse P2Y4 is closer to the rat ortholog in terms of agonist stimulation, while in terms of antagonist profile, the three P2Y4 receptor orthologs are similar.
Suarez-Huerta
N
N
Institut de Recherche Interdisciplinaire en Biologie Humaine et Nucléaire, Université Libre de Bruxelles, 808 route de Lennik, Bat. C 1070, Brussels, Belgium. nsuarez@ulb.ac.be
Pouillon
V
V
Boeynaems
J
J
Robaye
B
B
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D003001:Cloning, Molecular; D004305:Dose-Response Relationship, Drug; D006801:Humans; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology; D011732:Pyridoxal Phosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D051381:Rats; D018048:Receptors, Purinergic P2 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D013498:Suramin / Q000494:pharmacology; D014162:Transfection; D014227:Triazines / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014544:Uridine Triphosphate / Q000494:pharmacology
D000970:Antineoplastic Agents; D010975:Platelet Aggregation Inhibitors; D018048:Receptors, Purinergic P2; D011994:Recombinant Proteins; D014227:Triazines; C104749:purinoceptor P2Y4; C077792:pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid; C012842:Cibacron Blue F 3GA; D011732:Pyridoxal Phosphate; D013498:Suramin; D015544:Inositol 1,4,5-Trisphosphate; D000255:Adenosine Triphosphate; D014544:Uridine Triphosphate