Functional characterization of a basic D49 phospholipase A(2) (LmTX-I) from the venom of the snake Lachesis muta muta (bushmaster)

The whole venom of Lachesis muta muta is preponderantly neurotoxic but moderately myotoxic on the chick biventer cervicis preparation (BCp). We have now examined these toxic activities of a basic phospholipase A(2), LmTX-I, isolated from the whole venom. LmTX-I caused a significant concentration-dependent neuromuscular blockade in the BCp. The time to produce 50% neuromuscular blockade was 14.7 +/- 0.75 min (30 mu g/ml). 23.6 +/- 0.9 min (10 mu g/ml). 34 +/- 1.7 min (2.5 mu g/ml) and 39.2 +/- 3.6 min (1 mu g/ml), (n = 5/concentration; p < 0.05). Complete blockade with all tested concentrations was not accompanied by inhibition of the response to ACh. At the highest concentration. LmTX-I (30 mu g/ml) significantly reduced contractures elicited by exogenous KCl (20 mM), increased the release of creatine kinase (1542.5 +/- 183.9 IU[L vs 442.7 +/- 39.8 IU/L for controls after 120 min, p < 0.05), and induced the appearance of degenerating muscle fibers (similar to 15%). Quantification of myonecrosis indicated 14.8 +/- 0.8 and 2.0 +/- 0.4%, with 30 and 10 mu g/ml venom concentration, respectively, against 1.07 +/- 0.4% for control preparations. The findings indicate that the basic PLA(2) present on venom from L. m. muta (LmTX-1) possesses a dominant neurotoxic action on isolated chick nerve-muscle preparations, whereas myotoxicity was mainly observed at the highest Concentration used (30 mu g/ml). These effects of LmTX-I closely reproduce the effects of the whole venom of L. m. muta in chick neuromuscular preparations. (c) 2006 Elsevier Ltd. All rights reserved.47775976