Inhibition of phosphorylation of synapsin I and other synaptosomal proteins by beta-bungarotoxin, a phospholipase A2 neurotoxin.
Some snake venom neurotoxins, such as beta-bungarotoxin (beta-BuTX), which possess relatively low phospholipase A2 (PLA2) activity, act presynaptically to alter acetylcholine (ACh) release both in the periphery and in the CNS. In investigating the mechanism of this action, we found that beta-BuTX (5 and 15 nM) inhibited phosphorylation, in both resting and depolarized synaptosomes, of a wide range of proteins, including synapsin I. Naja naja atra PLA2, which has higher PLA2 activity, also inhibited phosphorylation but was less potent than beta-BuTX. At 1 nM, beta-BuTX and N. n. atra PLA2 inhibited phosphorylation of synapsin I only in depolarized synaptosomes. Synaptosomal ATP levels were not affected by 5 or 15 nM beta-BuTX or by 5 nM N. n. atra PLA2. Limited proteolysis, using Staphylococcus aureus V-8 protease, indicated that beta-BuTX inhibited phosphorylation of synapsin I in both the head and the tail regions. The inhibition of phosphorylation was not antagonized by nordihydroguaiaretic acid or indomethacin, suggesting that arachidonic acid derivatives do not mediate this inhibition. Furthermore, inhibition of phosphorylation by beta-BuTX and N. n. atra PLA2 was not altered in the presence of the phosphatase inhibitor okadaic acid, suggesting that stimulation of phosphatase activity is not responsible for this inhibition. Inhibition of protein phosphorylation by PLA2 neurotoxins and enzymes may be associated with an inhibition of ACh release.
Ueno
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E
Section of Pharmacology and Toxicology, School of Pharmacy, University of Connecticut, Storrs 06269.
Rosenberg
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P
D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; D000255:Adenosine Triphosphate / Q000032:analysis / Q000378:metabolism*; D000818:Animals; D002038:Bungarotoxins / Q000494:pharmacology*; D004546:Elapid Venoms / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004988:Ethers, Cyclic / Q000494:pharmacology; D006893:Hydroxyeicosatetraenoic Acids / Q000032:analysis / Q000378:metabolism*; D007213:Indomethacin / Q000494:pharmacology; D008297:Male; D009498:Neurotoxins / Q000494:pharmacology*; D019319:Okadaic Acid; D010741:Phospholipases A / Q000502:physiology; D054467:Phospholipases A2; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation / Q000187:drug effects; D011863:Radioimmunoassay; D051381:Rats; D017207:Rats, Sprague-Dawley; D012697:Serine Endopeptidases / Q000494:pharmacology; D013211:Staphylococcus aureus / Q000201:enzymology; D016704:Synapsins / Q000032:analysis / Q000378:metabolism*; D013574:Synaptosomes / Q000737:chemistry
D002038:Bungarotoxins; D004546:Elapid Venoms; D004988:Ethers, Cyclic; D006893:Hydroxyeicosatetraenoic Acids; D009498:Neurotoxins; D010761:Phosphorus Radioisotopes; D016704:Synapsins; D019319:Okadaic Acid; D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; D000255:Adenosine Triphosphate; D010741:Phospholipases A; D054467:Phospholipases A2; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; D007213:Indomethacin