title: Ligand-binding affinity of the type 1 and 2 inositol 1,4,5-trisphosphate receptors: effect of the membrane environment.
abstract: The inositol 1,4,5-trisphosphate (InsP3) receptor is essential for Ca2+ release from intracellular stores. There are three InsP3 receptor types which are targets for several types of regulation. Ca2+, phosphorylation, and protein-protein interactions may contribute to the complex pattern of the Ca2+ signal in stimulated cells. Furthermore, the 3 receptor types could have different affinities for InsP3. We compared the affinities of the type 1 receptor from the cerebellum with the liver type 2 receptor both in their membrane environment and after isolation by immunoprecipitation. Measurements of [3H]InsP3 binding in a cytosol-like medium revealed that the Kd of the liver receptor (45 +/- 5 nM, N = 14) was higher than the Kd of the cerebellar receptor (28 +/- 3 nM, N = 9). Solubilization and immunopurification of the liver InsP3 receptor resulted in a 10-fold increase in its affinity for InsP3. The affinity of the cerebellar receptor did not change under these conditions. Therefore, the extraction of the liver and the cerebellar receptors from their membrane environments induced an inversion of their relative affinities. Treatment of liver membranes with low concentrations of detergents also increased the affinity for InsP3 binding. These data indicate that the type 1 and the type 2 InsP3 receptors have different affinities for InsP3 and that the properties of the type 2 receptor are strongly regulated by hydrophobic interactions within its membrane environment.
authors.affiliation: INSERM U-442, Université Paris Sud, Orsay, France.
authors.forename: L
authors.initials: L
authors.lastname: Picard
authors.forename: J
authors.initials: J
authors.lastname: Ibarrondo
authors.forename: J F
authors.initials: JF
authors.lastname: Coquil
authors.forename: M
authors.initials: M
authors.lastname: Hilly
authors.forename: J P
authors.initials: JP
authors.lastname: Mauger
mesh_terms: D000818:Animals; D001667:Binding, Competitive; D015220:Calcium Channels / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D002531:Cerebellum / Q000166:cytology / Q000378:metabolism; D005260:Female; D066298:In Vitro Techniques; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism*; D053496:Inositol 1,4,5-Trisphosphate Receptors; D008024:Ligands; D008099:Liver / Q000166:cytology / Q000378:metabolism; D011233:Precipitin Tests; D051381:Rats; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D014316:Tritium
chemical_list: D015220:Calcium Channels; D053496:Inositol 1,4,5-Trisphosphate Receptors; D008024:Ligands; D018160:Receptors, Cytoplasmic and Nuclear; D014316:Tritium; D015544:Inositol 1,4,5-Trisphosphate