The coupling mechanism between endoplasmic reticulum (ER) calcium ion (Ca2+) stores and plasma membrane (PM) store-operated channels (SOCs) is crucial to Ca2+ signaling but has eluded detection. SOCs may be functionally related to the TRP family of receptor-operated channels. Direct comparison of endogenous SOCs with stably expressed TRP3 channels in human embryonic kidney (HEK293) cells revealed that TRP3 channels differ in being store independent. However, condensed cortical F-actin prevented activation of both SOC and TRP3 channels, which suggests that ER-PM interactions underlie coupling of both channels. A cell-permeant inhibitor of inositol trisphosphate receptor (InsP(2)R) function, 2-aminoethoxydiphenyl berate, prevented both receptor-induced TRP3 activation and store-induced SOC activation. It is concluded that InsP(3)Rs mediate both SOC and TRP channel opening and that the InsP(3)R is essential for maintaining coupling between store emptying and physiological activation of SOCs.