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Type: Artigo de periódico
Title: Purification and N-terminal sequencing of two presynaptic neurotoxic PLA(2), neuwieditoxin-I and neuwieditoxin-II,from Bothrops neuwiedi pauloensis (jararaca pintada) venom
Author: Borja-Oliveira, CR
Kassab, BH
Soares, AM
Toyama, MH
Giglio, JR
Marangoni, S
Re, L
Rodrigues-Simioni, L
Abstract: Two presynaptic phospholipases A(2) (PLA(2)), neuwieditoxin-I (NeuTX- I) and neuwieditoxin-II ( NeuTX-II), were isolated from the venom of Bothrops neuwiedi pauloensis (BNP). The venom was fractionated using molecular exclusion HPLC (Protein-Pak 300SW column), followed by reverse phase HPLC (mu Bondapak C18 column). Tricine-SDS-PAGE in the presence or absence of dithiothreitol showed that NeuTX- I and NeuTX-II had a molecular mass of approximately 14 kDa and 28kDa, respectively. At 10 mu g/ml, both toxins produced complete neuromuscular blockade in indirectly stimulated chick biventer cervicis isolated preparation without inhibiting the response to acetylcholine, but NeuTX-II reduced the response to KCl by 67.0 +/- 8.0% ( n= 3; p< 0.05). NeuTX- I and NeuTX-II are probably responsible for the presynaptic neurotoxicity of BNP venom in vitro. In fact, using loose patch clamp technique for mouse phrenic nerve- diaphragm preparation, NeuTX- I produced a calcium-dependent blockade of acetylcholine release and caused appearance of giant miniature end-plate potentials (mepps), indicating a pure presynaptic action. The N-terminal sequence of NeuTX- I was DLVQFGQMILKVAGRSLPKSYGAYGCYCGWGGRGK (71% homology with bothropstoxin-II and 54% homology with caudoxin) and that of NeuTX-II was SLFEFAKMILEETKRLPFPYYGAYGCYCGWGGQGQPKDAT (92% homology with Basp-III and 62% homology with crotoxin PLA2). The fact that NeuTX- I has Q-4 (Gln-4) and both toxins have F-5 (Phe-5) and Y-28 (Tyr-28) strongly suggests that NeuTX- I and NeuTX-II are Asp49 PLA2.
Subject: chick biventer cervicis
loose patch clamp
nerve-muscle preparation
neuromuscular junction
PLA(2) neurotoxin
presynaptic action
Bothrops neuwiedi pauloensis
Country: Brasil
Editor: Cevap-unesp
Citation: Journal Of Venomous Animals And Toxins Including Tropical Diseases. Cevap-unesp, v. 13, n. 1, n. 103, n. 121, 2007.
Rights: aberto
Identifier DOI: 10.1590/S1678-91992007000100008
Date Issue: 2007
Appears in Collections:Unicamp - Artigos e Outros Documentos

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