The inositol 1,4,5-trisphosphate receptor (InsP(3)R) is activated by InsP(3) binding to amino-terminal ligand binding domain (InsP(3)R-N). Recently we reported functional coupling of phosphatidylinositol (4, 5)-bisphosphate (PIP(2)) to the InsP(3)R. Specific binding of PIP(2) to InsP(3)R-N domain was postulated as a part of the InsP(3)R-PIP(2) functional coupling model. Here we utilized bacterially expressed and purified InsP(3)R-N domain to characterize its binding specificity for InsP(3), Adenophostin A (AdA) and the water-soluble PIP(2) analog dioctanoyl-(4,5)PIP(2) (ShPIP(2)). Obtained data led us to conclude that specific InsP(3), AdA, and ShPIP(2) binding sites are located within the InsP(3)R-N domain, that the extra receptor binding element responsible for enhanced binding of AdA is an integral part of the InsP(3)R-N domain, that ShPIP(2) is able to displace InsP(3) from the InsP(3)R-N, but InsP(3) or AdA is unable to completely displace ShPIP(2). These results support the InsP(3)R-PIP(2) functional coupling model and provide novel insights into InsP(3)R ligand specificity.
Copyright 2000 Academic Press.