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Activation of Early Events of the Mitogenic Response by a P2YPurinoceptor with Covalently Bound 3'-O-(4-Benzoyl)-Benzoyladenosine 5'-Triphosphate
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Activation of Early Events of the Mitogenic Response by a P2YPurinoceptor with Covalently Bound 3'-O-(4-Benzoyl)-Benzoyladenosine 5'-Triphosphate
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Activation of Early Events of the Mitogenic Response by a P2YPurinoceptor with Covalently Bound 3'-O-(4-Benzoyl)-Benzoyladenosine 5'-Triphosphate
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Journal Article
Activation of Early Events of the Mitogenic Response by a P2YPurinoceptor with Covalently Bound 3'-O-(4-Benzoyl)-Benzoyladenosine 5'-Triphosphate
1990
Overview
3'-O-(4-Benzoyl)benzoyl-ATP (BzATP), a photoaffinity analog of ATP, was used as a ligand for a P2Ypurinoceptor (adenine nucleotide receptor) present in intact Swiss 3T3 and 3T6 cells and A-431 epidermoid carcinoma cells. Photolysis of serum-starved cells in the presence of 10-50 μM BzATP, followed by extensive washing to remove unicorporated BzATP, induced the release of arachidonic acid. A trace (<0.01%) of photoincorporated BzATP was as effective as when 50 μM BzATP or ATP was contained in the incubation medium during the assay. Photoincorporated BzATP also stimulated the production of prostaglandin E2and the accumulation of cyclic AMP. In previous studies, we demonstrated that these three events are obligatory early steps in a pathway leading to DNA synthesis in the above cell lines. The evidence indicated that the purinoceptor activated by extracellular ATP or BzATP was linked to a pertussis toxin-sensitive GTP-binding protein. Consistent with these observations, we now find that pertussis toxin inhibits the effect of photoincorporated BzATP on arachidonic acid release. These results indicate that BzATP is an effective agonist for the P2Ypurinoceptor concerned with stimulation of DNA synthesis in 3T3, 3T6, and A-431 cells. Furthermore, after photolysis it becomes irreversibly associated with intact cells and promotes the activation of early events required for DNA synthesis.
