Antigenic, microbicidal and antiparasitic properties of an L-amino acid oxidase isolated from Bothrops jararaca snake venom

被引：88

作者：

Ciscotto, P. ^{[1
]}

de Avila, R. A. Machado ^{[1
]}

Coelho, E. A. F. ^{[1
]}

Oliveira, J. ^{[1
]}

Diniz, C. G. ^{[2
]}

Farias, L. M. ^{[2
]}

de Carvalho, M. A. R. ^{[2
]}

Maria, W. S. ^{[3
]}

Sanchez, E. F. ^{[3
]}

Borges, A. ^{[4
]}

Chavez-Olortegui, C. ^{[1
]}

机构：

[1] Univ Fed Minas Gerais, Inst Ciencias Biol, Dept Bioquim & Imunol, BR-30161970 Belo Horizonte, MG, Brazil

[2] Univ Fed Minas Gerais, Inst Ciencias Biol, Dept Microbiol, BR-30161970 Belo Horizonte, MG, Brazil

[3] Fundacao Ezequiel Dias, BR-30550010 Belo Horizonte, MG, Brazil

[4] Inst Estudios Avanzados, Ctr Biociencias & Med Mol, Lab Toxinas Anim, Caracas 1015A, Venezuela

来源：

TOXICON | 2009年 / 53卷 / 03期

关键词：

Bothrops jararaca; L-Amino acid oxidase; Microbicidal; Leishmanicidal; Neutralizing potency; APOPTOSIS-INDUCING FACTOR; HUMAN PLATELET-AGGREGATION; NAJA-KAOUTHIA VENOM; FUNCTIONAL-CHARACTERIZATION; ANTIBOTHROPIC ANTIVENOM; NEUTRALIZING POTENCY; OXIDATIVE STRESS; APOXIN I; PURIFICATION; ANTIBACTERIAL;

D O I：

10.1016/j.toxicon.2008.12.004

中图分类号：

R9 [药学];

学科分类号：

100702 [药剂学];

摘要：

Venoms from the bee Apis mellifera, the caterpillar Lonomia achelous, the spiders Lycosa sp. and Phoneutria nigriventer, the scorpions Tityus bahiensis and Tityus serrulatus, and the snakes Bothrops alternates, Bothrops jararaca, Bothrops jararacussu, Bothrops moojeni, Bothrops neuwiedi, Crotalus durissus terrificus, and Lachesis muta were assayed (800 mu g/mL) for activity against Staphylococcus aureus. Venoms from B. jararaca and B. jararacussu showed the highest S. aureus growth inhibition and also against other Gram-positive and Gram-negative bacteria. To characterize the microbicidal component(s) produced by B. jararaca, venom was fractionated through gel exclusion chromatography. The high molecular weight, anti-S. aureus P1 fraction was further resolved by anion exchange chromatography through Mono Q columns using a 0-0.5 M NaCl gradient. Bactericidal Mono Q fractions P5 and P6 showed significant LAAO activity using L-leucine as substrate. These fractions were pooled and subjected to Heparin affinity chromatography, which rendered a single LAAO activity peak. The anti-S. aureus activity was abolished by catalase, suggesting that the effect is dependent on H2O2 production. SDS-PAGE of isolated LAAO indicated the presence of three isoforms since deglycosylation with a recombinant N-glycanase rendered a single 38.2 kDa component. B. jararaca LAAO specific activity was 142.7 U/mg, based on the oxidation of L-leucine. The correlation between in vivo neutralization of lethal toxicity (ED50) and levels of horse therapeutic antibodies anti-LAAO measured by ELISA was investigated to predict the potency of Brazilian antibothropic antivenoms. Six horses were hyperimmunized with Bothrops venoms (50% from B. jararaca and 12.5% each from B. alternatus, B. jararacussu, B. neuwiedii and B. moojeni). To set up an indirect ELISA, B. jararaca LAAO and crude venom were used as antigens. Correlation coefficients (r) between ED50 and ELISA antibody titers against B. jararaca venom and LAAO were 0.846 (p < 0.001) and 0.747 (p < 0.001), respectively. The hemolytic and leishmanicidal (anti-Leishmania amazonensis) activity of LAAO was also determined. (C) 2008 Published by Elsevier Ltd.

引用

页码：330 / 341

页数：12

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