Differential effects of snake venom phospholipase A2 neurotoxin (beta-bungarotoxin) and enzyme (Naja naja atra) on protein kinases.
The phospholipase A2 (PLA2) neurotoxin, beta-bungarotoxin (beta-BuTX), presynaptically alters acetylcholine release. We previously found that beta-BuTX inhibits protein phosphorylation in rat brain synaptosomes. This inhibition was not due to the inhibition of ATP synthesis, the action of arachidonic acid (AA) metabolites, or the stimulation of phosphatase activities. A typical PLA2 enzyme from Naja naja atra (N. n. atra) venom also inhibited phosphorylation but with lesser potency than that of beta-BuTX. We now report the effects of beta-BuTX and N. n. atra PLA2 on the activities of protein kinases. Treatments of synaptic plasma membrane or cytosol with N. n. atra PLA2 stimulated the activities of cAMP-dependent kinase, Ca2+/calmodulin-dependent kinase II, and protein kinase C (PKC), whereas beta-BuTX had no effect on these kinases. Calyculin A, a phosphatase-1 and -2A inhibitor, increased the stimulation of phosphorylation by N. n. atra PLA2, indicating that the stimulation is not due to an inhibition of phosphatase activities. The stimulation of PKC by N. n. atra PLA2 appears to be mediated by free fatty acids (FFAs) resulting from phospholipid hydrolysis by PLA2, since (1) treatment of either synaptic plasma membrane or cytosol with N. n. atra PLA2 produced large amounts of FFAs, and (2) AA, an exogenous FFA, stimulated PKC activity to an extent similar to that caused by N. n. atra PLA2. Thus, the mechanisms of action of beta-BuTX and N. n. atra PLA2 appear quite different from each other although both agents inhibit phosphorylation in intact synaptosomes.
Ueno
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E
Department of Pharmaceutical Sciences, University of Connecticut, School of Pharmacy, Storrs 06269, USA.
Rosenberg
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P
D000818:Animals; D001921:Brain / Q000187:drug effects / Q000201:enzymology; D002038:Bungarotoxins / Q000633:toxicity*; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D003600:Cytosol / Q000201:enzymology; D017815:Elapidae; D005230:Fatty Acids, Nonesterified / Q000096:biosynthesis; D066298:In Vitro Techniques; D008297:Male; D010741:Phospholipases A / Q000633:toxicity*; D054467:Phospholipases A2; D011493:Protein Kinase C / Q000378:metabolism; D011494:Protein Kinases / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012910:Snake Venoms / Q000201:enzymology* / Q000633:toxicity*; D013570:Synaptic Membranes / Q000201:enzymology; D013574:Synaptosomes / Q000187:drug effects / Q000201:enzymology
D002038:Bungarotoxins; D005230:Fatty Acids, Nonesterified; D012910:Snake Venoms; D011494:Protein Kinases; D017868:Cyclic AMP-Dependent Protein Kinases; D011493:Protein Kinase C; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D010741:Phospholipases A; D054467:Phospholipases A2