[32P]3'-O-(4-benzoyl)benzoyl ATP as a photoaffinity label for a phospholipase C-coupled P2Y-purinergic receptor.
作者： J.L. Boyer , C.L. Cooper , T.K. Harden
DOI: 10.1016/S0021-9258(18)77377-8
关键词:
摘要: A 32P-labelled ATP analog, 3'-O-(4-benzoyl)benzoyl (BzATP) previously shown to be an agonist at P2Y-purinergic receptors (Boyer J. L., and Harden T. K. (1989) Mol. Pharmacol. 36, 831-835), has been used as a probe for the receptor on turkey erythrocyte plasma membranes. In absence of light, [32P]BzATP bound membranes with high affinity (KD approximately 5 nM), in saturable reversible manner. The binding was competitively inhibited by ADP analogs (2-methylthioadenosine 5'-triphosphate greater than adenosine 5'-O-(2-thiodiphosphate) BzATP beta,gamma-methyleneadenosine 5'-adenylylimidodiphosphate) pharmacological specificity consistent that receptor. Guanine nucleotides (guanosine 5'-O-(3-thiotriphosphate) GTP guanosine GMP) noncompetitively radioligand. Photolysis [32P] BzATP-prelabeled resulted incorporation radiolabel into protein 53,000 Da. Photolabeling concentration-dependent manner potency order characteristic modulated guanine nucleotides. daltons also labeled from several other tissues known express These results suggest can label covalently this radioprobe will useful reagent further characterization purification
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