Basic Phospholipase A2 Research Articles

Identification of domains of phosphatidylcholine in human erythrocyte plasma membranes. Differential action of acidic and basic phospholipases A2 from Agkistrodon halys blomhoffii.

Highly purified acidic (pI 4.9) and basic (pI 8.7) phospholipases A2 from snake (Agkistrodon halys blomhoffii) venom hydrolyzed approximately 20% and 60%, respectively, of the phosphatidylcholine (PC) of intact human erythrocytes prior to hemolysis. Sequential use of the acidic enzyme followed by the basic phospholipase A2 or vice versa manifested a characteristic PC hydrolysis pattern. For example, when acidic enzyme had hydrolyzed nearly 20% of this substrate, a subsequent treatment with the basic enzyme hydrolyzed only an additional 40% of the PC before hemolysis. On the other hand, in experiments where hydrolysis of about 20% of PC of erythrocytes was achieved by a short term incubation with the basic enzyme, then a further treatment of the same cells with the acidic enzyme caused only 10% additional PC hydrolysis before hemolysis. This demonstrated that the acidic enzyme hydrolyzed one domain of PC in the intact erythrocytes, whereas the basic enzyme hydrolyzed not only the same but also another domain of PC in membranes. Analysis of fatty acids released by the action of these two phospholipases A2 on erythrocytes indicated further characteristic differences. In particular, the ratio of released saturated to unsaturated fatty acids was significantly higher with the acidic enzyme as compared with the basic phospholipase A2. These results provide firm support to the conclusion that there are different domains of PC in human erythrocyte membranes and that the acidic and basic phospholipase A2 of A. halys blomhoffii can be used to identify them.

Postsynaptic effects of crotoxin and of its isolated subunits.

Crotoxin is a potent neurotoxin from the venom of Crotalus durissus terrificus. It is composed of two subunits: a basic phospholipase A2 with low toxicity (component B) and an acidic protein seemingly devoid of intrinsic biological activity (component A). Crotoxin and its isolated phospholipase subunit block the depolarisation caused by cholinergic agonists on the isolated electroplaque from Electrophorus electricus. The other component, which is inactive when applied alone, enhances the pharmacological activity of the phospholipase when the two components are used together. Crotoxin also blocks the increase of 22Na+ efflux caused by carbamylcholine from excitable microsacs prepared from Torpedo marmorata electric organ. Crotoxin therefore acts postsynaptically, but does not interfere with the binding of alpha-toxin from Naja nigricollis to the nicotinic cholinergic receptor site. Instead, like local anesthetics, it stabilizes a desensitized form of the acetylcholine receptor characterized by its high affinity for agonists. The phospholipase component B binds in a non-saturable manner to receptor-rich membranes. In contrast, component A does not bind to acetylcholine receptor-rich membranes, but completely prevents the non-saturable binding of component B. When the two components are applied together, a saturable binding of the latter is observed with the acetylcholine receptor-rich membranes.