Inositol 1,4,5-triphosphate (Ins(1,4,5)P3) is a second messenger that regulates Ca2+ channels in many important cell signalling pathways. In sea urchin sperm the outer investment of the egg triggers the acrosome reaction (AR) that involves Ins(1,4,5)P3 production and the opening of two Ca2+ channels. Here we have sought to identify a high-affinity Ins(1,4,5)P3 receptor in Strongylocentrotus purpuratus sperm. An Ins(1,4,5)P3 binding component was affinity-purified 12-fold from sperm extracts. It displayed similar characteristics to the Ins(1,4,5)P3 receptor from other sources: pH-dependent high affinity for Ins(1,4,5)P3 (KD = 261 nM), a tau1/2 of association and dissociation of 50 and 40 s, respectively, specificity (IC50 > 5 microM for Ins(1)P1, Ins(1,4)P2 and Ins(1,3,4,5)P4), and pharmacological sensitivity (10 and 100 microg heparin/ml inhibited 75% and 100% binding respectively). An antibody against the carboxy-terminal of the type I Ins(1,4,5)P3 receptor of somatic cells recognised a plasma membrane component in the sperm head and less intensely in the flagella. This antibody also recognised a 240 kDa band from isolated head plasma membranes, and weakly in flagellar membrane. This IP3 receptor-like protein may mediate the sustained uptake of Ca2+ through the second Ca2+ channel opened during the AR.