title: Interaction of polyanions with the recognition sites for inositol 1,4,5-trisphosphate in the bovine adrenal cortex.
abstract: Inositol 1,4,5-trisphosphate (InsP3) serves as a second messenger for Ca2+ mobilization in a wide variety of cells. InsP3 activates a specific receptor/channel located on an internal Ca2+ store. Because heparin has already been shown to block the action of InsP3, we have looked at the influence of other polyanions (dextran sulfate and polyvinyl sulfate) on the action and metabolism of InsP3 in the bovine adrenal cortex. Polyvinyl sulfate blocked InsP3 binding to adrenal cortex microsomes with a half-maximal efficiency of 250 nM. Scatchard analyses revealed that this effect was not competitive. The Ca2+ releasing activity of InsP3 on the same microsomal preparation was monitored with the fluorescent indicator, fura-2. Polyvinyl sulfate blocked this activity with a half-maximal efficiency of 80 nM. The effect of polyvinyl sulfate could not be overcome by supramaximal doses of InsP3, suggesting a non-competitive inhibitory effect. The activity of InsP3 phosphatase from bovine adrenal cortex microsomes was also studied. Polyvinyl sulfate inhibited the activity of the phosphatase with a half-maximal efficiency of 5 microM. Lineweaver-Burk plots revealed that this effect was not competitive. Polyvinyl sulfate was able to inhibit the activity of InsP3 kinase from bovine adrenal cortex cytosol. The half-maximal dose was 15 nM and the Lineweaver-Burk analysis showed that the inhibition was not competitive. The effect of dextran sulfate 5000 (DS-5000) on these activities was also studied. DS-5000 inhibited in a competitive manner the binding of InsP3 to its receptor (IC50 of 34 microM), the release of Ca2+ induced by InsP3 (IC50 of 6.5 microM) and the activity of InsP3 phosphatase (IC50 of 57 microM).(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Pharmacology, Faculty of Medicine, University of Sherbrooke, Quebec, Canada.
authors.forename: T T
authors.initials: TT
authors.lastname: Luong
authors.forename: M
authors.initials: M
authors.lastname: Poitras
authors.forename: G
authors.initials: G
authors.lastname: Boulay
authors.forename: G
authors.initials: G
authors.lastname: Guillemette
mesh_terms: D000302:Adrenal Cortex / Q000187:drug effects / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism; D002417:Cattle; D016264:Dextran Sulfate / Q000494:pharmacology*; D006493:Heparin / Q000494:pharmacology; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism*; D007700:Kinetics; D015636:Magnesium Chloride / Q000494:pharmacology; D008861:Microsomes / Q000187:drug effects / Q000378:metabolism; D010770:Phosphotransferases / Q000037:antagonists & inhibitors / Q000187:drug effects / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor); D011145:Polyvinyls / Q000494:pharmacology*
chemical_list: D011145:Polyvinyls; D015636:Magnesium Chloride; C019050:polyvinyl sulfate; D015544:Inositol 1,4,5-Trisphosphate; D006493:Heparin; D016264:Dextran Sulfate; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C051038:Inositol 1,4,5-trisphosphate 3-kinase; D002118:Calcium