A phospholipase A2-stimulating protein regulated by protein kinase C in Aplysia neurons.
We describe some properties on an Mr 30,000 thermolabile and trypsin-sensitive protein that activates phospholipase A2 (PLA2) and which was isolated from nervous tissue of the marine mollusk, Aplysia californica. A similar protein is present in rat cerebral cortex. This protein was partially purified from crude homogenates of nervous tissue by ion exchange chromatography on DEAE-Sephadex followed by size-exclusion high performance liquid chromatography (HPLC). It is loosely associated with membrane fractions, and is extracted by 0.05% Tween 20. Although similar in size to several previously described PLA2-stimulating proteins from non-neural mammalian cells and tissues, it differs from them in some aspects of biological activity. The protein promotes the release of eicosanoids from the membranes of intact Aplysia neurons prelabeled with [3H]arachidonic acid and appears to be an in vitro substrate for protein kinase C (PKC). PLA2-stimulating activity is greatly enhanced after exposing isolated ganglia to phorbol dibutyrate (PDBu) and is reduced by treatment with immobilized E. coli alkaline phosphatase. These observations suggest that phosphorylation of this stimulatory protein by PKC regulates PLA2 in neurons.
Calignano
A
A
Howard Hughes Medical Institute, Center for Neurobiology and Behavior, Columbia University, New York, NY 10032.
Piomelli
D
D
Sacktor
T C
TC
Schwartz
J H
JH
D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; D000818:Animals; D001048:Aplysia; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D005724:Ganglia / Q000378:metabolism*; D006893:Hydroxyeicosatetraenoic Acids / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D008970:Molecular Weight; D009474:Neurons / Q000378:metabolism*; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D011493:Protein Kinase C / Q000378:metabolism*; D011506:Proteins / Q000302:isolation & purification / Q000378:metabolism*; D014357:Trypsin
D006893:Hydroxyeicosatetraenoic Acids; D011506:Proteins; C055324:phospholipase A2-activating protein; D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; D011493:Protein Kinase C; D010741:Phospholipases A; D054467:Phospholipases A2; D014357:Trypsin