Pancreastatin (PST) receptors have been recently shown to mediate activation of phospholipase C (PLC) in rat liver membranes. There is evidence that the G protein that links pancreastatin receptor with PLC-beta is pertussis toxin-insensitive and belongs to the G(alpha)q family. Here, we have employed blocking antisera to sort out the specific PLC-beta isoform as well as the specific G(alpha) subunit activated by PST receptor in rat liver membranes. The presence of different PLC-beta isoforms was checked by immunoblot analysis. Only PLC-beta4 was not detected, whereas PLC-beta1, beta2 and beta3 were abundant in rat liver membranes. However, only anti-PLC-beta3 serum was able to block the PST receptor response. We also checked the expression of G(alpha)q and Galpha11 in rat liver membranes by immunoblot. Even though both isoforms were present. only anti-Galpha11 serum was able to block the PST receptor response. In order to check the specificity of the blocking antisera, we employed them to block the effect of ADP and thrombin stimulating PLC activity in platelet membranes, a system lacking Galpha11. Anti-G(alpha)q but not anti-Galpha11 sera were able to block the agonist stimulated PLC activity. These data suggest that PST receptor response is mediated by the activation of the beta3 isoform of PLC via Galpha11 protein stimulation in rat liver membranes.