Inhibition of rat parotid ecto-ATPase activity

The inhibitory profile of several known and suspected ecto-ATPase inhibitors was compared on ecto-ATPase activity in rat parotid plasma membranes. Those chemicals with high IC50 (above 130 μM) were the nucleotides α,β-methylene ATP, β,γ-methylene ATP, 2-methylthio ATP, inosine triphosphate, 5′-p-fluorosulphonylbenzoyladenosine, the sulphonates, 1-amino-2-naphthol-4-sulphonic acid, Coomassie brilliant blue G, and the stilbene disulphonates, DIDS and SITS. Those agents with low IC50 were: Coomassie brilliant blue R (114 μM), ATPγS (49 μM), suramin (72 μM) and Reactive blue 2 (28 μM). The last three inhibitors have similar potencies as inhibitors of ATP hydrolysis by whole parotid acinar cells. ARL67156, a selective inhibitor of ecto-ATPase, had an IC50 of approx. 120 μM. Suramin displayed non-competitive inhibition of ecto-ATPase whereas the inhibitory effects of ATPγS and Reactive blue 2 were curvilinear on Dixon plots. These results define the effects of various agents on ecto-ATPase in an exocrine tissue that has been shown to respond to extracellular ATP.