Different localization of inositol 1,4,5-trisphosphate and ryanodine binding sites in rat liver

Abstract

The distribution of inositol 1,4,5-trisphosphate and ryanodine binding sites between plasma membrane, microsomal, and mitochondrial fractions of rat liver were compared. IP 3 bound mostly to the plasma membrane fraction (K d = 6 nM; B max = 802 fmol/mg protein). Some IP 3 binding sites were also present in the microsomal and mitochondrial fractions (K d = 2.5 and 2.9 nM; B max = 35 and 23 fmol/mg protein respectively). The possibility that these binding sites are due to contamination of the fractions with plasma membrane cannot be excluded. Binding of IP 3 to the plasma membrane was inhibited by heparin but not by either caffeine or tetracaine. High-affinity ryanodine binding sites were present mostly in the microsomal fraction (K d = 13 nM; B max = 301 fmol/mg protein). Lower affinity binding sites were also found to be present in the mitochondrial and plasma membrane fractions. Binding of ryanodine to the microsomal fraction was inhibited by both caffeine and tetracaine but not by heparin. These data demonstrate that IP 3 and ryanodine binding sites are present in different cellular compartments in the liver. These differences in the localization of the binding sites might be indicative of their functional differences.