Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/198320
Type: Artigo de periódico
Title: Neuromuscular Activity Of Batx, A Presynaptic Basic Pla2 Isolated From Bothrops Alternatus Snake Venom.
Author: Ponce-Soto, L A
Barros, J C
Marangoni, S
Hernandez, S
Dal Belo, C A
Corrado, A P
Hyslop, S
Rodrigues-Simioni, L
Abstract: We have previously isolated a Lys49 phospholipase A(2) homolog (BaTX) from Bothrops alternatus snake venom using a combination of molecular exclusion chromatography and reverse phase HPLC and shown its ability to cause neuromuscular blockade. In this work, we describe a one-step procedure for the purification of this toxin and provide further details of its neuromuscular activity. The toxin was purified by reverse phase HPLC and its purity and molecular mass were confirmed by SDS-PAGE, MALDI-TOF mass spectrometry, amino acid analysis and N-terminal sequencing. BaTX (0.007-1.4 microM) produced time-dependent, irreversible neuromuscular blockade in isolated mouse phrenic nerve-diaphragm and chick biventer cervicis preparations (time to 50% blockade with 0.35 microM toxin: 58+/-4 and 24+/-1 min, respectively; n=3-8; mean+/-S.E.) without significantly affecting the response to direct muscle stimulation. In chick preparations, contractures to exogenous acetylcholine (55 and 110 microM) or KCl (13.4 mM) were unaltered after complete blockade by all toxin concentrations. These results, which strongly suggested a presynaptic mechanism of action for this toxin, were reinforced by (1) the inability of BaTX to interfere with the carbachol-induced depolarization of the resting membrane, (2) a significant decrease in the frequency and amplitude of miniature end-plate potentials, and (3) a significant reduction (59+/-4%, n=12) in the quantal content of the end-plate potentials after a 60 min incubation with the toxin (1.4 microM). In addition, a decrease in the organ bath temperature from 37 degrees C to 24 degrees C and/or the replacement of calcium with strontium prevented the neuromuscular blockade, indicating a temperature-dependent effect possibly mediated by enzymatic activity.
Subject: Animals
Bothrops
Calcium
Chick Embryo
Cholinergic Agonists
Chromatography, High Pressure Liquid
Crotalid Venoms
Diaphragm
Dose-response Relationship, Drug
Electric Stimulation
Electrophoresis, Polyacrylamide Gel
Male
Mice
Miniature Postsynaptic Potentials
Molecular Weight
Neuromuscular Blocking Agents
Neuromuscular Junction
Phospholipases A2
Phrenic Nerve
Presynaptic Terminals
Sequence Analysis, Protein
Spectrometry, Mass, Matrix-assisted Laser Desorption-ionization
Synaptic Transmission
Temperature
Time Factors
Citation: Comparative Biochemistry And Physiology. Toxicology & Pharmacology : Cbp. v. 150, n. 2, p. 291-7, 2009-Aug.
Rights: fechado
Identifier DOI: 10.1016/j.cbpc.2009.05.007
Address: http://www.ncbi.nlm.nih.gov/pubmed/19463969
Date Issue: 2009
Appears in Collections:Unicamp - Artigos e Outros Documentos

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