FEBS Letters | |
Characterization of myocardial extracellular ATP receptors by photoaffinity labelling and functional assays | |
Powers, Kit1  Scarpa, Antonio1  Giannattasio, Bartolo1  | |
[1] Department of Physiology and Biophysics, Case Western Reserve University, 2119 Abington Rd., Cleveland, OH 44116, USA | |
关键词: ATP receptor; Photolabelling; Myocyte; 8-Azido-ATP; ATPγS; adenosine-5′-O-(3-thiotriphosphate); 2Az-ATP; 2-azido-adenosine-5′-triphosphate; 8Az-ATP; 8-azido-adenosine-5′-triphosphate; Bz-ATP; 3′-O-(4-benzoyl)benzoyl-[α-32P]adenosine-5′-triphosphate; BSA; bovine serum albumin; 2-meSATP; 2-methylthio- adenosine-5′-triphosphate; fura2/AM; fura2-acetoxymethyl ester; GTP; guanosine-5′-triphosphate; SDS; sodium dodecyl sulfate; UTP; uridine-5′-triphosphate; | |
DOI : 10.1016/0014-5793(92)80582-2 | |
学科分类:生物化学/生物物理 | |
来源: John Wiley & Sons Ltd. | |
【 摘 要 】
Extracellular ATP receptors in rat ventricular myocytes were investigated through intact cell photolabelling followed by protein isolation. 8-Azido-ATP (8Az-ATP) was used for labelling under specific conditions determined by parallel functional studies. In those studies ATP-induced cytosolic Ca2+ transients were irreversibly and specifically inhibited by UV-photolyzed 8Az-ATP, but not by 2-azido-ATP (2Az-ATP), even in the presence of high concentrations of phosphonucleotides not affecting myocardial ATP receptors. Under those conditions background labelling is minimized and radioactive 8Az-ATP specifically labels a band of 45–48 kDa on a SDS gel. Labelling under the above conditions in the presence of ATPγS or 2-methylthio-ATP (2-meSATP), which are distinct for two functionally different cardiac ATP receptors, shows two different proteins within the same band consistent with the possible labelling of these two receptors.
【 授权许可】
Unknown
【 预 览 】
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