title: Exposure of phosphatidylcholine and phosphatidylinositol in plasma membranes from rat brain synaptosomes treated with phospholipase A2 toxins (beta-bungarotoxin, notexin) and enzymes (Naja nigricollis, Naja naja atra).
abstract: Phospholipase A2 (PLA2) toxins act presynaptically to block acetylcholine release and are much more potent and specific in their actions than PLA2 enzymes even though they have lower enzymatic activity. Since their mechanism of action is not completely understood, it was of interest to examine the toxins' effects on phospholipid asymmetry as changes in asymmetry are associated with changes in membrane functioning. Rat brain synaptosomes were treated with the PLA2 toxins beta-bungarotoxin (beta-BuTx) and notexin and with the PLA2 enzymes Naja nigricollis and Naja naja atra under relatively non-disruptive conditions as judged by leakage of lactate dehydrogenase (LDH) and levels of phospholipid hydrolysis. The exposure of phosphatidylcholine (PC) and phosphatidylinositol (PI) on the synaptosomal surface was investigated by means of a specific PC-exchange protein (PCEP) and a PI-specific phospholipase C (PI-PLC), respectively. Treatment of the synaptosomes with N. nigricollis PLA2, beta-BuTx and notexin did not affect the availability of PC to exchange by PCEP, but significantly increased the exposure of PI to hydrolysis by PI-PLC. In contrast, N. n. atra PLA2 slightly decreased the exposure of PC and did not affect that of PI. The differences between N. n. atra PLA2, on the one hand, and N. nigricollis PLA2, beta-BuTx and notexin, on the other hand, parallel differences in their pharmacological activities. Our earlier studies showed that PLA2 enzymes, and possibly PLA2 toxins, have a pharmacological site separate from the enzymatic site. Since in the present study the effect on PI was abolished by EDTA, the presence of an enzymatic site in addition to the pharmacological site may be required or alternatively divalent cations may be required for the effects on PI asymmetry independent of the inhibition of PLA2 by EDTA.
authors.affiliation: Basil and Gerald Felsenstein Medical Research Center, Petah Tikva, Israel.
authors.forename: R
authors.initials: R
authors.lastname: Shina
authors.forename: R C
authors.initials: RC
authors.lastname: Crain
authors.forename: P
authors.initials: P
authors.lastname: Rosenberg
authors.forename: E
authors.initials: E
authors.lastname: Condrea
mesh_terms: D000818:Animals; D001923:Brain Chemistry; D002038:Bungarotoxins / Q000494:pharmacology; D002462:Cell Membrane / Q000737:chemistry; D004546:Elapid Venoms / Q000201:enzymology / Q000494:pharmacology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008563:Membrane Lipids / Q000032:analysis*; D010713:Phosphatidylcholines / Q000032:analysis*; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D010716:Phosphatidylinositols / Q000032:analysis*; D054801:Phosphoinositide Phospholipase C; D010741:Phospholipases A / Q000187:drug effects / Q000378:metabolism*; D054467:Phospholipases A2; D010727:Phosphoric Diester Hydrolases; D051381:Rats; D013574:Synaptosomes / Q000737:chemistry*
chemical_list: D002038:Bungarotoxins; D004546:Elapid Venoms; D008563:Membrane Lipids; D010713:Phosphatidylcholines; D010716:Phosphatidylinositols; C005082:notexin; D007770:L-Lactate Dehydrogenase; D010741:Phospholipases A; D054467:Phospholipases A2; D010727:Phosphoric Diester Hydrolases; D054801:Phosphoinositide Phospholipase C; D043265:Phosphatidylinositol Diacylglycerol-Lyase