Guanine nucleotide and pyrophosphate activate exogenous phosphatidylinositol 4,5-bisphosphate hydrolysis in rat liver plasma membranes.
5'-guanylylimidodiphosphate (GppNHp) in the presence of deoxycholate, stimulated the phospholipase C-mediated hydrolysis of exogenous [3H]phosphatidylinositol 4,5-bisphosphate ([3H]PIP2) to myo-[3H]inositol 1,4,5-trisphosphate in rat liver plasma membranes. Activation was not specific for guanine nucleotides as 5'-adenylylimidodiphosphate, imidodiphosphate and pyrophosphate stimulated the enzyme with similar efficacies and potencies. Enzyme activation by GppNHp was most pronounced when [3H]PIP2 was used as substrate. No added Ca++ was required for [3H]PIP2 breakdown but hydrolysis was inhibited by divalent ion chelators. GppNHp stimulation was apparent in the presence of Ca++ or Mg++ as well as chelator concentrations that partially inhibited the enzyme, indicating that this effect was not attributed to changes in affinity of these divalent cations for the enzyme or substrate. These results suggest that guanine nucleotides can stimulate the hydrolysis of exogenous [3H]PIP2 in rat liver membranes by a non-specific effect probably due to the interaction of the diphosphate moiety with the enzyme or substrate.
Bojanic
D
D
Department of Biochemistry, University of Tennessee, Memphis 38163.
Wallace
M A
MA
Wojcikiewicz
R J
RJ
Fain
J N
JN
D002118:Calcium / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism*; D003840:Deoxycholic Acid / Q000494:pharmacology; D011756:Diphosphates / Q000502:physiology*; D019204:GTP-Binding Proteins / Q000502:physiology*; D006150:Guanine Nucleotides / Q000502:physiology*; D006165:Guanylyl Imidodiphosphate / Q000494:pharmacology; D006868:Hydrolysis; D008099:Liver / Q000378:metabolism*; D008274:Magnesium / Q000378:metabolism; D010716:Phosphatidylinositols / Q000378:metabolism*; D010738:Type C Phospholipases / Q000378:metabolism*
D011756:Diphosphates; D006150:Guanine Nucleotides; D010716:Phosphatidylinositols; D003840:Deoxycholic Acid; D006165:Guanylyl Imidodiphosphate; D010738:Type C Phospholipases; D019204:GTP-Binding Proteins; D008274:Magnesium; D002118:Calcium