Rat parietal cells were incubated for 2 h with pertussis toxin (100 ng/ml) which ADP-ribosylates and inactivates guanine nucleotide regulatory proteins (G proteins) of the 'Gi-like' family. The effect of this pretreatment on the action of inhibitors of parietal cell acid secretion was investigated by using the accumulation of the weak base aminopyrine as an index of secretory activity. The inhibitory actions of near maximally effective concentrations of prostaglandin E2 (PGE2), somatostatin and epidermal growth factor (EGF) on histamine-stimulated aminopyrine accumulation were reduced by 83%, 72% and 70%, respectively, by preincubation with pertussis toxin. By contrast, the inhibitory action of a near maximally effective concentration of 12-O-tetradecanoylphorbol 13-acetate on histamine-stimulated aminopyrine accumulation was reduced by only 12%. It is concluded that G-proteins are involved in the inhibitory actions of PGE2, somatostatin and EGF on parietal cells. However, since the inhibitory actions of PGE2 and EGF can be distinguished by the blockade of the action of EGF, but not that of PGE2, by 3-isobutyl-1-methylxanthine, it is possible that PGE2 and EGF either activate the same G-protein in different ways or work through different G-proteins.