| http://purl.uniprot.org/citations/14742685 | http://www.w3.org/1999/02/22-rdf-syntax-ns#type | http://purl.uniprot.org/core/Journal_Citation |
| http://purl.uniprot.org/citations/14742685 | http://www.w3.org/2000/01/rdf-schema#comment | "The human P2Y1 receptor (P2Y1-R) was purified after high-level expression from a recombinant baculovirus in Sf9 insect cells. Quantification by protein staining and with a radioligand binding assay using the high-affinity P2Y1-R antagonist [3H]MRS2279 ([3H]2-chloro-N6-methyl-(N)-methanocarba-2'-deoxyadenosine 3',5'-bis-phosphate) indicated a nearly homogenous preparation of receptor protein. Ki values determined in [3H]MRS2279 binding assays for antagonists with the purified P2Y1-R were in good agreement with the Ki and KB values determined for these molecules in membrane binding and activity assays, respectively. Availability of P2Y1-R in purified form allowed direct determination of nucleotide agonist affinities under conditions not compromised by nucleotide metabolism/interconversion, and an order of affinities of 2-methylthio-ADP (2MeSADP) > ADP = 2-methylthioATP = adenosine-5'-O-(3-thio)triphosphate = adenosine-5'-O(2-thiodiphosphate) >> ATP was obtained. The signaling activity of the purified P2Y1-R was quantified after reconstitution in proteoliposomes with heterotrimeric G proteins. Steady-state GTP hydrolysis in vesicles reconstituted with P2Y1-R and Galpha(q)beta(1)gamma(2) was stimulated by the addition of either 2MeADP or RGS4 alone and was increased by up to 50-fold in their combined presence. EC50 values of agonists for activation of the purified P2Y1-R were similar to their respective Ki values determined in radioligand binding experiments with the purified receptor. Moreover, ATP exhibited 20-fold higher EC50 and Ki values than did ADP and was a partial agonist relative to ADP and 2MeSADP under conditions in which no metabolism of the nucleotide occurred. Both RGS4 and PLC-beta1 were potent and efficacious GTPase-activating proteins for Galphaq and Galpha11 in P2Y1-R-containing vesicles. These results illustrate that the binding and signaling properties of the human P2Y1-R can be studied with purified proteins under conditions that circumvent the complications that occur in vivo."xsd:string |
| http://purl.uniprot.org/citations/14742685 | http://purl.org/dc/terms/identifier | "doi:10.1124/mol.65.2.426"xsd:string |
| http://purl.uniprot.org/citations/14742685 | http://purl.uniprot.org/core/author | "Harden T.K."xsd:string |
| http://purl.uniprot.org/citations/14742685 | http://purl.uniprot.org/core/author | "Waldo G.L."xsd:string |
| http://purl.uniprot.org/citations/14742685 | http://purl.uniprot.org/core/date | "2004"xsd:gYear |
| http://purl.uniprot.org/citations/14742685 | http://purl.uniprot.org/core/name | "Mol Pharmacol"xsd:string |
| http://purl.uniprot.org/citations/14742685 | http://purl.uniprot.org/core/pages | "426-436"xsd:string |
| http://purl.uniprot.org/citations/14742685 | http://purl.uniprot.org/core/title | "Agonist binding and Gq-stimulating activities of the purified human P2Y1 receptor."xsd:string |
| http://purl.uniprot.org/citations/14742685 | http://purl.uniprot.org/core/volume | "65"xsd:string |
| http://purl.uniprot.org/citations/14742685 | http://www.w3.org/2004/02/skos/core#exactMatch | http://purl.uniprot.org/pubmed/14742685 |
| http://purl.uniprot.org/citations/14742685 | http://xmlns.com/foaf/0.1/primaryTopicOf | https://pubmed.ncbi.nlm.nih.gov/14742685 |
| http://purl.uniprot.org/uniprot/#_O14775-mappedCitation-14742685 | http://www.w3.org/1999/02/22-rdf-syntax-ns#object | http://purl.uniprot.org/citations/14742685 |
| http://purl.uniprot.org/uniprot/#_P50148-mappedCitation-14742685 | http://www.w3.org/1999/02/22-rdf-syntax-ns#object | http://purl.uniprot.org/citations/14742685 |
| http://purl.uniprot.org/uniprot/#_H3BLF7-mappedCitation-14742685 | http://www.w3.org/1999/02/22-rdf-syntax-ns#object | http://purl.uniprot.org/citations/14742685 |
| http://purl.uniprot.org/uniprot/#_P16520-mappedCitation-14742685 | http://www.w3.org/1999/02/22-rdf-syntax-ns#object | http://purl.uniprot.org/citations/14742685 |
| http://purl.uniprot.org/uniprot/#_Q3U1B1-mappedCitation-14742685 | http://www.w3.org/1999/02/22-rdf-syntax-ns#object | http://purl.uniprot.org/citations/14742685 |
| http://purl.uniprot.org/uniprot/#_H3BKR2-mappedCitation-14742685 | http://www.w3.org/1999/02/22-rdf-syntax-ns#object | http://purl.uniprot.org/citations/14742685 |
| http://purl.uniprot.org/uniprot/#_P50150-mappedCitation-14742685 | http://www.w3.org/1999/02/22-rdf-syntax-ns#object | http://purl.uniprot.org/citations/14742685 |
| http://purl.uniprot.org/uniprot/#_P50151-mappedCitation-14742685 | http://www.w3.org/1999/02/22-rdf-syntax-ns#object | http://purl.uniprot.org/citations/14742685 |
| http://purl.uniprot.org/uniprot/#_Q3UHH5-mappedCitation-14742685 | http://www.w3.org/1999/02/22-rdf-syntax-ns#object | http://purl.uniprot.org/citations/14742685 |
| http://purl.uniprot.org/uniprot/#_O14610-mappedCitation-14742685 | http://www.w3.org/1999/02/22-rdf-syntax-ns#object | http://purl.uniprot.org/citations/14742685 |
| http://purl.uniprot.org/uniprot/#_O60262-mappedCitation-14742685 | http://www.w3.org/1999/02/22-rdf-syntax-ns#object | http://purl.uniprot.org/citations/14742685 |
| http://purl.uniprot.org/uniprot/#_O95837-mappedCitation-14742685 | http://www.w3.org/1999/02/22-rdf-syntax-ns#object | http://purl.uniprot.org/citations/14742685 |
| http://purl.uniprot.org/uniprot/#_P29992-mappedCitation-14742685 | http://www.w3.org/1999/02/22-rdf-syntax-ns#object | http://purl.uniprot.org/citations/14742685 |