A Ca(2+)-independent activation of a type IV cytosolic phospholipase A(2) underlies the receptor stimulation of arachidonic acid-dependent noncapacitative calcium entry.
The oscillatory [Ca(2+)](i) signals typically seen following physiologically relevant stimulation of phospholipase C-linked receptors are associated with a receptor-activated entry of Ca(2+), which plays a critical role in driving the oscillations and influencing their frequency. We have recently shown that this receptor-activated entry of Ca(2+) does not conform to the widely accepted "capacitative" model and, instead, reflects the activity of a distinct, novel Ca(2+) entry pathway regulated by arachidonic acid (Shuttleworth, T. J., and Thompson, J. L. (1998) J. Biol. Chem. 273, 32636-32643). We now show that the generation of arachidonic acid under these conditions results from the activity of a type IV cytosolic phospholipase A(2) (cPLA(2)). Although cPLA(2) activation commonly involves a Ca(2+)-dependent translocation to the membrane, at these low agonist concentrations cPLA(2) activation was independent of increases in [Ca(2+)](i), and no detectable translocation to the membrane occurs. Nevertheless, stimulation of cPLA(2) activity was confined to the membrane fraction, where an increase in phosphorylation of the enzyme was observed. We suggest that, at the low agonist concentrations associated with oscillatory [Ca(2+)](i) signals, cPLA(2) activation involves an increased phosphorylation of a discrete pool of the total cellular cPLA(2) that is already localized within the membrane fraction at resting [Ca(2+)](i).
Osterhout
J L
JL
Department of Pharmacology and Physiology, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642, USA.
Shuttleworth
T J
TJ
D016718:Arachidonic Acid / Q000378:metabolism*; D001095:Arachidonic Acids / Q000494:pharmacology; D001692:Biological Transport; D000440:Butanols / Q000494:pharmacology; D020013:Calcium Signaling; D002217:Carbachol / Q000494:pharmacology*; D002451:Cell Compartmentation; D002460:Cell Line; D003600:Cytosol / Q000201:enzymology; D004553:Electric Conductivity; D004789:Enzyme Activation; D000431:Ethanol / Q000494:pharmacology; D054513:Group IV Phospholipases A2; D006801:Humans; D018721:Muscarinic Agonists / Q000494:pharmacology; D009281:Naphthalenes / Q000494:pharmacology; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D010741:Phospholipases A / Q000145:classification / Q000187:drug effects / Q000378:metabolism*; D011753:Pyrones / Q000494:pharmacology; D043587:Receptor, Muscarinic M3; D011976:Receptors, Muscarinic / Q000378:metabolism*
D001095:Arachidonic Acids; D000440:Butanols; D018721:Muscarinic Agonists; D009281:Naphthalenes; D010726:Phosphodiesterase Inhibitors; D011753:Pyrones; D043587:Receptor, Muscarinic M3; D011976:Receptors, Muscarinic; C081565:arachidonyltrifluoromethane; D016718:Arachidonic Acid; D000431:Ethanol; C068314:6-(bromomethylene)tetrahydro-3-(1-naphthaleneyl)-2H-pyran-2-one; D002217:Carbachol; D010741:Phospholipases A; D054513:Group IV Phospholipases A2; C516389:PLA2G4A protein, human