Interaction of benzene 1,2,4-trisphosphate with inositol 1,4,5-trisphosphate receptor and metabolizing enzymes.
In a wide variety of cells, inositol 1,4,5-trisphosphate (InsP3) is an important second messenger involved in the regulation of intracellular Ca2+ concentration. InsP3 interacts with specific receptors and triggers the release of sequestered Ca2+ from an internal store. We have synthesized a structural analogue of InsP3 by phosphorylation of the free hydroxyl groups of 1,2,4-benzenetriol with dibenzylphosphorochloridate. The product benzene 1,2,4-trisphosphate (BzP3) was shown to interact with InsP3 receptor and InsP3 metabolizing enzymes of bovine adrenal cortex. BzP3 competitively blocked InsP3 binding to adrenal cortex microsomes with a half-maximal efficiency at 34 microM. This affinity was about 10,000 times lower than that of InsP3 for its receptor. The Ca2+ releasing activity of BzP3 on the same microsomal preparation was monitored with the fluorescent indicator fura-2. BzP3 had no agonistic effect on this activity but it was able to inhibit InsP3-induced Ca2+ release in a dose-dependent manner. The activity of InsP3 phosphatase was also studied. BzP3 inhibited the activity of the phosphatase with a half-maximal efficiency of 32 microM. BzP3 was also able to inhibit the activity of the cytosolic InsP3 kinase with a half-maximal efficiency of 6.1 microM. These results show that BzP3 is interacting with the three specific recognition sites for InsP3 in the bovine adrenal cortex. The inhibitory effect of this compound is relatively more potent on the metabolizing enzymes than on the Ca(2+)-mobilizing receptor.
Department of Pharmacology, Faculty of Medicine, University of Sherbrooke, Quebec, Canada.
M
M
Poitras
S
S
Bernier
G
G
Boulay
A
A
Fournier
G
G
Guillemette
D000311:Adrenal Glands / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels; D002417:Cattle; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism* / Q000494:pharmacology; D053496:Inositol 1,4,5-Trisphosphate Receptors; D008861:Microsomes / Q000378:metabolism; D010755:Organophosphates / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology*; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*; D010766:Phosphorylation; D010770:Phosphotransferases / Q000378:metabolism*; D017853:Phosphotransferases (Alcohol Group Acceptor); D011956:Receptors, Cell Surface / Q000378:metabolism*; D018160:Receptors, Cytoplasmic and Nuclear
D015220:Calcium Channels; D053496:Inositol 1,4,5-Trisphosphate Receptors; D010755:Organophosphates; D011956:Receptors, Cell Surface; D018160:Receptors, Cytoplasmic and Nuclear; C080376:benzene 1,2,4-trisphosphate; D015544:Inositol 1,4,5-Trisphosphate; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C051038:Inositol 1,4,5-trisphosphate 3-kinase; D010744:Phosphoric Monoester Hydrolases; C043422:myo-inositol 1,4,5-triphosphate phosphatase; D002118:Calcium