Inositol hexakisphosphate (InsP6) stimulates 45Ca2+ influx in purified mitochondria from rat liver. The action of InsP6 is concentration-dependent, with an apparent EC50 value of 50 microM. Stimulation of 45Ca2+ influx may follow the interaction between InsP6 and specific membrane receptors. Accordingly, [3H]InsP6 binds to specific and saturable recognition sites in purified mitochondria. These results support the view that InsP6 acts as a signal molecule to regulate the intracellular homeostasis of Ca2+.