Categories
Articles

Acta Biochim Biophys Sin 2004,36(12): Purification and Characterization of Jararassin-I, A Thrombin-like Enzyme from Bothrops jararaca Snake Venom

  • Post author By abbs

https://www.abbs.info     
E-mail: [email protected]

ISSN
1672-9145                                              
 Acta Biochim Biophys Sin
2004, 36(12): 798–802                                                
 CN 31-1940/Q

Purification and Characterization of Jararassin-I, A Thrombin-like
Enzyme from Bothrops jararaca Snake Venom

Débora F. VIEIRA1, Leandra WATANABE1, Carolina
D. SANT´ANA2,3, Silvana MARCUSSI2,4, Suely V. SAMPAIO3,
Andreimar M. SOARES2*, and Raghuvir K. ARNI1*

1Departamento
de Física, Instituto de Biociências, Letras e Ciências Exatas, Universidade
Estadual Paulista,

UNESP, 15054-000, São José do Rio Preto, SP, Brazil;

2Unidade de
Biotecnologia, Universidade de Ribeirão Preto, UNAERP, Ribeirão Preto, SP,
Brazil;

3Departamento
de Análises Clínicas, Toxicológicas e Bromatológicas, FCFRP, Universidade de
São Paulo, USP, Ribeirão Preto, SP, Brazil;

4Departamento
de Bioquímica e Imunologia, FMRP, Universidade de São Paulo, USP, Ribeirão
Preto, SP, Brazil

Abstract        A thrombin-like serine protease, jararassin-I, was isolated from the
venom of Bothrops jararaca. The protein was obtained in high yield and
purity by a single chromatographic step using the affinity resin Benzamidine-Sepharose
CL-6B. SDS-PAGE and dynamic light scattering analyses indicated that the
molecular mass of the enzyme was about 30 kD. The enzyme possessed
fibrinogenolytic and coagulant activities. The jararassin-I degraded the Bb chain of
fibrinogen while the Aa chain and g chain were unchanged. Proteases inhibitors, PMSF and benzamidine
inhibited the coagulant activity. These results showed jararassin-I is a serine
protease similar to coagulating thrombin-like snake venom proteases, but it
specifically cleaves Bb chain of bovine fibrinogen. Single crystals of enzyme were obtained
(0.2 mm×0.2 mm×0.2 mm) and used for X-ray diffraction experiments.

Key words        snake venom; Bothrops jararaca; serine protease thrombin-like;
fibrinogenolytic activity; crystallization

 

—————–

Received: September 22, 2004        Accepted: October 29,
2004

Abbreviations: SVTLE, snake venom thrombin-like enzyme; SDS, sodium
dodecyl sulphate; EDTA, ethylene diaminetetraacetic acid

*Corresponding authors:

Andreimar M. SOARES: Tel, 55-16-603-6892;
Fax, 55-16-603-7030; E-mail, [email protected] 

Raghuvir K. ARNI: E-mail, [email protected]