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Purification and characterization of a hyaluronidase from venom of the spider Vitalius dubius (Araneae, Theraphosidae)
Sutti, Rafael; Tamascia, Mariana Leite; Hyslop, Stephen; Rocha-e-Silva, Thomaz Augusto Alves.
Afiliação
  • Sutti, Rafael; Santa Casa de São Paulo. Medical School. Department of Physiological Sciences. São Paulo. BR
  • Tamascia, Mariana Leite; State University of Campinas. School of Medical Science. Department of Pharmacology. Campinas. BR
  • Hyslop, Stephen; Federal University of Campinas. School of Medical Sciences. Department of Pharmacology. Campinas. BR
  • Rocha-e-Silva, Thomaz Augusto Alves; Santa Casa de Sao Paulo. Medical School. Department of Physiological Sciences. São Paulo. BR
J. venom. anim. toxins incl. trop. dis ; J. venom. anim. toxins incl. trop. dis;20: 1-7, 04/02/2014. graf
Article em En | LILACS | ID: lil-702582
Biblioteca responsável: BR33.1
ABSTRACT

Background:

Venom hyaluronidase (Hyase) contributes to the diffusion of venom from the inoculation site. In this work, we purified and characterized Hyase from the venom of Vitalius dubius (Araneae, Theraphosidae), a large theraphosid found in southeastern Brazil. Venom obtained by electrical stimulation of adult male and female V. dubius was initially fractionated by gel filtration on a Superdex® 75 column. Active fractions were pooled and applied to a heparin-sepharose affinity column. The proteins were eluted with a linear NaCl gradient.

Results:

Active fractions were pooled and assessed for purity by SDS-PAGE and RP-HPLC. The physicochemical tests included optimum pH, heat stability, presence of isoforms, neutralization by flavonoids and assessment of commercial antivenoms. Hyase was purified and presented a specific activity of 148 turbidity-reducing units (TRU)/mg (venom 36 TRU/mg; purification factor of ~4). Hyase displayed a molecular mass of 43 kDa by SDS-PAGE. Zymography in hyaluronic-acid-containing gels indicated an absence of enzyme isoforms. The optimum pH was 4-5, with highest activity at 37°C. Hyase was stable up to 60°C; but its activity was lost at higher temperatures and maintained after several freeze-thaw cycles. The NaCl concentration (up to 1 M) did not influence activity. Hyase had greater action towards hyaluronic acid compared to chondroitin sulfate, and was completely neutralized by polyvalent antiarachnid sera, but not by caterpillar, scorpion or snakes antivenoms.

Conclusion:

The neutralization by arachnid but not scorpion antivenom indicates that this enzyme shares antigenic epitopes with similar enzymes in other spider venoms. The biochemical properties of this Hyase are comparable to others described.
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Texto completo: 1 Coleções: 01-internacional Base de dados: LILACS Assunto principal: Picada de Aranha / Venenos de Aranha Limite: Animals Idioma: En Revista: J. venom. anim. toxins incl. trop. dis Assunto da revista: TOXICOLOGIA Ano de publicação: 2014 Tipo de documento: Article País de afiliação: Brasil País de publicação: Brasil
Texto completo
Adicionar na Minha BVS
Imprimir
XML
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: LILACS Assunto principal: Picada de Aranha / Venenos de Aranha Limite: Animals Idioma: En Revista: J. venom. anim. toxins incl. trop. dis Assunto da revista: TOXICOLOGIA Ano de publicação: 2014 Tipo de documento: Article País de afiliação: Brasil País de publicação: Brasil