Requirement of adenosine 5'-triphosphate and Ca2+ for guanosine 5'-triphosphate-binding protein-mediated phospholipase D activation in rat pheochromocytoma PC12 cells.
In digitonin-permeabilized PC12 cells labeled with [3H]palmitic acid, formation of [3H]phosphatidylethanol ([3H]PEt), a marker of phospholipase D (PLD) activity, was increased with increasing concentrations of Ca2+ in the presence of ATP, Mg2+ and ethanol. Guanosine-5'-O-(3-thio-triphosphate) (GTP gamma S) significantly enhanced Ca(2+)-stimulated [3H]PEt formation. The effect of GTP gamma S was abolished when any one of Ca2+, ATP or Mg2+ was excluded. ATP could not be replaced by other nucleotides except for ADP. Thus, the GTP-binding protein-mediated PLD activation absolutely required both Ca2+ and Mg-ATP.
Department of Biochemistry, Gifu University School of Medicine, Japan.
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Kanoh
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Kanaho
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Nozawa
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000502:physiology*; D002463:Cell Membrane Permeability / Q000187:drug effects; D000242:Cyclic AMP / Q000502:physiology*; D004789:Enzyme Activation / Q000187:drug effects; D000431:Ethanol / Q000494:pharmacology; D019204:GTP-Binding Proteins / Q000378:metabolism / Q000502:physiology*; D020404:Glycerophospholipids; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D008566:Membranes / Q000187:drug effects / Q000201:enzymology; D016716:PC12 Cells; D010712:Phosphatidic Acids / Q000378:metabolism; D010739:Phospholipase D / Q000378:metabolism*; D051381:Rats
D020404:Glycerophospholipids; D010712:Phosphatidic Acids; C051521:phosphatidylethanol; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D000431:Ethanol; D000255:Adenosine Triphosphate; D000242:Cyclic AMP; D010739:Phospholipase D; D019204:GTP-Binding Proteins; D002118:Calcium