PROTEOLYTIC ACTIVITY OF AMAZONIAN SCORPION Tityus metuendus VENOM (SCORPIONES: BUTHIDAE)

ARAÚJO T. F. (1, 3), HIGA A. M. (1, 3), NORONHA  M. D. N. (1, 3), MARTINS M. J. P. (2), MUNIZ E. G. (3), MEDEIROS B. M. (4), AGUIAR N. O. (4), LOPEZ-LOZANO J. L. (1,3).

(1) Programa de Pós-Graduação em Biotecnologia e Recursos Naturais – Universidade do Estado do Amazonas; (2) Programa de Pós-Graduação em Biotecnologia – Universidade Federal do Amazonas; (3) Fundação de Medicina Tropical do Amazonas; (4) Curso de Ciências Biológicas da Universidade Federal do Amazonas.

Neurotoxin peptides are the main constituents of scorpion venoms. Few studies about Amazonian scorpion neurotoxins have been reported. Enzymes studies of scorpion’s venoms are necessaries to better understand of biological function this venom. Tityus metuendus specimens are principal responsible of scorpionism in Manaus region. The aim of this study was research in Tityus metuendus venom toxins with phospholipasic and proteolytic activities. The scorpions were captured in Manaus region (Amazonas State – Brazil) and the venom was obtained by low voltage electric stimulation of telson venom gland. Pool sample venom (90 g) was submitted to SDS – PAGE reduction or non reduction conditions to detected 14kDa proteins constituents. The phospholipasic activity was tested using 3% egg yolk incorporated in 1% agarose plate gel method in buffer PBS 0,10 mM pH 8,1 and 0,09 mM CaCl2. Proteolytic zimogram activity profile was obtained using different substrates (casein, hemoglobin, gelatin, and fibrinogen) in buffer solution pH 6. 5 – 8. 5 ranges. Gelatin and casein substrates were degraded by proteolytic activity of venom using glicine 100 mM pH 8. 3 buffer. The zimogram profile suggests gelatinolytic and caseinolytic enzymes with 23 kDa and 50 kDa respectively. These enzymes were inhibited by serine protease inhibitors. A Phospholipasic activity was not detected. Purification of proteolytic enzymes are in curse to know structure / biological function relationship.

KEY WORD: scorpion, Tityus metuendus, serine proteases, Amazonia.

FINANCIAL SUPPORT: CNPQ, FAPEAM, FMTAM, FINEP.

CORREPONDENSE TO:THIAGO FERREIRA DE ARAÚJO, Centro de Ofidismo Prof. Paulo Friedrich Bürheim, Fundação de Medicina Tropical do Amazonas, Manaus, Brasil. Phone: + 55 92 3238 42 94. E-mail: thiago_biologo@hotmail.com.