Regulation of inositol 1,4,5-trisphosphate receptors in rat basophilic leukemia cells. I. Multiple conformational states of the receptor in a microsomal preparation.

A detailed characterization of the inositol 1,4,5-trisphosphate (IP3) receptor in rat basophilic leukemia (RBL) cells, a neoplastic mast cell line, has been possible through the growth of solid RBL cell tumors which provide a rich source of IP3 receptor. Equilibrium binding studies show a 1.6 +/- 0.1 pmol/mg of protein maximal binding capacity for [3H]IP3 at optimal Ca2+ (10 microM). The specificity of the RBL cell IP3 receptor towards phosphoinositides, ATP and heparin parallels those previously described with excitable and nonexcitable tissues. [3H]IP3 binding is slightly enhanced from < 1 nM to 10 microM Ca2+ and inhibited by > 10 microM Ca2+. Kinetic and equilibrium studies provide evidence for at least two classes or conformational states of binding sites with pico- and nanomolar affinities. At nM concentrations of IP3, neither binding to the IP3 receptor nor IP3-induced Ca2+ efflux from permeabilized cells demonstrates cooperativity. In contrast, at pM concentrations, IP3 binding kinetics deviate from simple mass action suggesting a complex interaction among binding sites for IP3 on the receptor-channel oligomer. The mechanisms that regulate [3H]IP3 binding in RBL cells are unique when compared to what has been reported in other cells.