Biochemical and biological evaluation of gyroxin isolated from Crotalus durissus terrificus venom

Author(s):Show less -

Barros, L. C. ^{[1, 2]} ; Soares, A. M. ^[3] ; Costa, F. L. ^[4] ; Rodrigues, V. M. ^[4] ; Fuly, A. L. ^[5] ; Giglio, J. R. ^[6] ; Gallacci, M. ^[7] ; Thomazini-Santos, I. A. ^[2] ; Barraviera, S. R. C. S. ^{[1, 8]} ; Barraviera, B. ^{[1, 2]} ; Ferreira Junior, R. S. ^{[1, 2]}

Affiliation

^[1] UNESP, Sao Paulo State Univ, CEVAP, BR-18618000 Botucatu, SP - Brazil

^[2] UNESP, Sao Paulo State Univ, Botucatu Med Sch, Dept Trop Dis & Imaging Diag, BR-18618000 Botucatu, SP - Brazil

^[3] Univ Sao Paulo, Dept Clin Toxicol & Bromatol Anal, Sch Pharmaceut Sci, BR-14049 Ribeirao Preto, SP - Brazil

^[4] Univ Fed Uberlandia, Inst Genet & Biochem, BR-38400 Uberlandia, MG - Brazil

^[5] Univ Fed Fluminense, Inst Biol, Dept Cellular & Mol Biol, Niteroi, RJ - Brazil

^[6] Univ Sao Paulo, Med Sch Ribeirao Preto, Dept Biochem & Immunol, Ribeirao Preto, SP - Brazil

^[7] UNESP, Sao Paulo State Univ, Inst Biol, Dept Pharmacol, BR-18618000 Botucatu, SP - Brazil

^[8] UNESP, Sao Paulo State Univ, Botucatu Med Sch, Dept Dermatol, BR-18618000 Botucatu, SP - Brazil

• Document type: Journal article
• Source: Journal of Venomous Animals and Toxins including Tropical Diseases; v. 17, n. 1, p. 23-33, 2011.
• Web of Science Citations: 32

Abstract

Gyroxin, a thrombin-like enzyme isolated from Crotalus durissus terrificus venom and capable of converting fibrinogen into fibrin, presents coagulant and neurotoxic activities. The aim of the present study was to evaluate such coagulant and toxic properties. Gyroxin was isolated using only two chromatographic steps - namely gel filtration (Sephadex G-75) and affinity (Benzamidine Sepharose 6B) - resulting in a sample of high purity, as evaluated by RP-HPLC C2/C18 and electrophoretic analysis that showed a molecular mass of 30 kDa. Gyroxin hydrolyzed specific chromogenic substrates, which caused it to be classified as a serine proteinase and thrombin-like enzyme. It was stable from pH 5.5 to 8.5 and inhibited by Mn²+, Cu²+, PMSF and benzamidine. Human plasma coagulation was more efficient at pH 6.0. An in vivo toxicity test showed that only behavioral alterations occurred, with no barrel rotation. Gyroxin was not able to block neuromuscular contraction in vitro, which suggests that its action, at the studied concentrations, has no effect on the peripheral nervous system. (AU)

• FAPESP's process: 07/05159-7 - Isolation of coagulant serine-proteases from Bothrops neuwiedi pauloensis and Crotalus durissus terrificus venoms: functional and structural characterization
• Grantee: Benedito Barraviera
• Support Opportunities: Regular Research Grants