Interaction of (3H)arylazido aminopropionyl ATP ((3H)ANAPP3) with P2- aurinergic receptors in the smooth muscle of the isolated guinea-pig vas deferens.
Fedan-JS; Hogaboom-GK; O'Donnell-JP; Jeng-SJ; Guillory-RJ
Eur J Pharmacol 1985 Jan; 108(1):49-61
Experiments were begun with tritium labeled arylazido-aminopropionyl- ATP (ANAPP3) for the purpose of understanding the biochemical nature of the P2-purinergic receptor. Intact vasa deferentia were treated with ANAPP3 and samples of homogenate, cytosol and a crude membrane fraction were analyzed using sodium-dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Photolysis of ANAPP3 resulted in the incorporation of radioactivity into cellular components with apparent molecular weights of 54 to 66 and 43 to 57 kilodaltons. Photoincorporation of ANAPP3 was associated with the crude membrane fraction and not the cytosol, was reduced in the presence of ATP in an ATP concentration dependent manner, was lessened following pretreatment of the tissues with photolyzed nonradiolabeled ANAPP3, and was not affected by the nucleoside transport inhibitor, dipyridamole. Tension studies resulted in a concentration dependent reduction in the initial contractile response to ANAPP3. The response to tritium was antagonized in tissues which had been pretreated with nonradiolabeled ANAPP3. Dipyridamole had no effect on the contractile response to ANAPP3. The authors suggest that the antagonism by photolyzed ANAPP3 of adenine nucleotide induced responses was a direct result of the covalent insertion at or near the recognition site of cell surface P2-purinergic receptors.
NIOSH-Author; Reproductive-hazards; Laboratory-animals; Reproductive-system-disorders; Cytotoxic-effects; In-vitro-studies; Cell-damage; Nucleosides
European Journal of Pharmacology