Palytoxin (PTX) is known to bind to Na,K-ATPase, to inhibit its activity, and to induce cation conductance, but the mechanism of these effects is still poorly understood. In Xenopus oocytes, PTX induced a large cation conductance, an effect that could be prevented or reversed by ouabain for oocytes expressing Xenopus Na,K-pumps but not with those expressing Bufo Na,K-pumps. In both cases patch-clamp experiments demonstrated a 7-8 pS channel in the presence of PTX. A large PTX-induced conductance could be observed with minimal Na,K-pump inhibition. From the single PTX-induced channel and macroscopic whole oocyte conductance, and the number of Na,K-pumps, we can conclude that PTX-induced conductance occurs through a direct interaction of PTX with a small number of Na,K-pumps.