Purification and characterization of jararassin-I, A thrombin-like enzyme from Bothrops jararaca snake venom.
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 x 0.2 mm x 0.2 mm) and used for X-ray diffraction experiments.
Vieira
Dëora F
DF
Departamento de Física, Instituto de Biociências Exatas, Universidade Estadual Paulista, UNESP, São José do Rio Preto, SP, Brazil.
Watanabe
Leandra
L
Sant'ana
Carolina D
CD
Marcussi
Silvana
S
Sampaio
Suely V
SV
Soares
Andreimar M
AM
Arni
Raghuvir K
RK
D000818:Animals; D001777:Blood Coagulation / Q000187:drug effects; D017837:Bothrops; D002846:Chromatography, Affinity; D003435:Crotalid Venoms / Q000201:enzymology*; D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D005340:Fibrinogen / Q000378:metabolism; D012697:Serine Endopeptidases / Q000235:genetics* / Q000378:metabolism*; D014757:Viper Venoms / Q000235:genetics* / Q000378:metabolism*
D003435:Crotalid Venoms; D014757:Viper Venoms; D005340:Fibrinogen; D012697:Serine Endopeptidases; C502623:jararassin-I, Bothops jararaca