Identificação, purificação e determinação da estrutura e função de componentes de baixas massas moleculares do do veneno da aranha Phoneutria nigriventer (Araneae; Ctenidae)
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Data
2009-08-06
Autores
Gomes, Paulo César [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
As aranhas do gênero Phoneutria (Aranae, Ctenidae) são comumente conhecidas como aranha armadeira ou aranha da banana, devido à posição de ataque/defesa que elas assumem contra uma presa/competidor e devido sua alta incidência nas plantações de bananas. Estas aranhas são solitárias, errantes (não constroem teias), muito agressivas e responsáveis por muitos casos graves de acidentes por envenenamento e de morte registrados. Estas aranhas são amplamente distribuídas nas regiões temperadas da América do Sul, com várias espécies já descritas. A espécie Phoneutria nigriventer é mais comum nas regiões centroeste e sudeste do Brasil. Um grande número de neurotoxinas tem sido purificado de venenos de aranhas do gênero Phoneutria. No entanto, as neurotoxinas não – protéicas de baixa massa molecular precisam ser isoladas e caracterizadas estrutural e funcionalmente. Essas toxinas são de potencial interesse na neuroquímica como ferramentas para investigações do sistema nervoso. O objetivo do presente estudo foi a caracterização estrutural e funcional da Nigriventrina, uma nova neurotoxina não-protéica de baixa massa molecular, isolada da fração hidrofílica do veneno da aranha Phoneutria nigriventer por (RPHPLC) sob gradiente de acetonitrila. A elucidação estrutural foi realizada com HRESIMS, ESI-MS, ESI-MS/MS e espectroscopia de Ressonância Magnética Nuclear 1D e 2D. A toxina apresentou massa molecular de 422Da (C12H14N4O13) e foi caracterizada como hidrazil-dioxopiperidina. A caracterização biológica foi realizada pela aplicação icv da Nigriventrina em cérebro de rato, seguido pelo acompanhamento da expressão da proteína Fos e dupla marcação imunoistoquímica. Todos os neurônios duplamente marcados foram associados ao receptor ionotrópico de glutamato NMDA (N-metil-D-aspartato), subtipo NMDA-NR1. A Nigriventrina apresentou afinidade...
The spiders of the genus Phoneutria (Aranae, Ctenidae) are commonly known as “armed spider” or “banana spider”, because of the attack–defense position that they assume against a prey/competitor and their high incidence in banana plantations. These spiders are solitary, wandering (no web), very aggressive, responsible for many cases of severe envenomation and recorded mortality. These spiders are widely distributed in the warm regions of South America, and several species have been described. The Phoneutria nigriventer are the most common in central and south-eastern regions of Brazil. A large number of neurotoxins have been purified from the venoms of Phoneutria spiders. However, the non-proteic low molecular mass neurotoxins must be isolated and structural and functionally characterized. These toxins have an interesting potential in neurochemistry as tools for investigations of nervous system. The objective of the present study was the structural and functional characterization of “Nigriventrine”, a novel non-proteic low molecular mass neurotoxin, isolated from the hydrophilic fraction of Phoneutria nigriventer spider venom by (RP-HPLC) under gradient of acetonitrile. The structural elucidation was carried out with HRESIMS, ESI-MS, ESI-MS/MS and Nuclear Magnetic Resonance 1D and 2D spectroscopy. The toxin presented molecular mass of 422Da (C12H14N4O13) and was characterized as hydroxyl-hydrazyl-dioxopiperidine. The biological characterization was performed by the i.c.v application of nigriventrine in rat brain, followed by the monitoring of the expression of Fos protein and doublelabeling immunohistochemistry. All doubly labeled neurons were associated to Nmethyl- D- aspartate/subtype of ionotropic glutamate receptor (NMDA-NR1). The nigriventrine presented affinity mainly to cortical regions, which are linked to perceptions and the voluntary muscles control of the animals... (Complete abstract click electronic access below)
The spiders of the genus Phoneutria (Aranae, Ctenidae) are commonly known as “armed spider” or “banana spider”, because of the attack–defense position that they assume against a prey/competitor and their high incidence in banana plantations. These spiders are solitary, wandering (no web), very aggressive, responsible for many cases of severe envenomation and recorded mortality. These spiders are widely distributed in the warm regions of South America, and several species have been described. The Phoneutria nigriventer are the most common in central and south-eastern regions of Brazil. A large number of neurotoxins have been purified from the venoms of Phoneutria spiders. However, the non-proteic low molecular mass neurotoxins must be isolated and structural and functionally characterized. These toxins have an interesting potential in neurochemistry as tools for investigations of nervous system. The objective of the present study was the structural and functional characterization of “Nigriventrine”, a novel non-proteic low molecular mass neurotoxin, isolated from the hydrophilic fraction of Phoneutria nigriventer spider venom by (RP-HPLC) under gradient of acetonitrile. The structural elucidation was carried out with HRESIMS, ESI-MS, ESI-MS/MS and Nuclear Magnetic Resonance 1D and 2D spectroscopy. The toxin presented molecular mass of 422Da (C12H14N4O13) and was characterized as hydroxyl-hydrazyl-dioxopiperidine. The biological characterization was performed by the i.c.v application of nigriventrine in rat brain, followed by the monitoring of the expression of Fos protein and doublelabeling immunohistochemistry. All doubly labeled neurons were associated to Nmethyl- D- aspartate/subtype of ionotropic glutamate receptor (NMDA-NR1). The nigriventrine presented affinity mainly to cortical regions, which are linked to perceptions and the voluntary muscles control of the animals... (Complete abstract click electronic access below)
Descrição
Palavras-chave
Aracnideo, Aranha - Veneno, Toxinas, Neuroquimica, Toxinas de baixa massa molecular, Spider venoms, Low molecular mass toxins, Neurochemistry
Como citar
GOMES, Paulo César. Identificação, purificação e determinação da estrutura e função de componentes de baixas massas moleculares do do veneno da aranha Phoneutria nigriventer (Araneae; Ctenidae). 2009. 89 f. Tese (doutorado) - Universidade Estadual Paulista, Instituto de Biociências de Rio Claro, 2009.