Open AccessEnzymological characterization of FII a , a fibrinolytic enzyme from Agkistrodon acutus venom 1
Author(s) -
LIANG Xiuxia,
ZHOU Yingna,
CHEN Jiashu,
QIU Pengxin,
CHEN Huizhen,
SUN Huanhuan,
WU Yuping,
YAN Guangmei
Publication year - 2005
Publication title -
acta pharmacologica sinica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.514
H-Index - 90
eISSN - 1745-7254
pISSN - 1671-4083
DOI - 10.1111/j.1745-7254.2005.00204.x
Subject(s) - enzyme , biology , medicine , chemistry , biochemistry
Aim: To study the enzymological characterization of a fibrinolytic enzyme (FII a ) from Agkistrodon acutus venom. Methods: The fibrinogenolytic effect and the influences of several protease inhibitors, chelating agents, and metal ions on fibrinogenolytic activity were visualized by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis. The metal content of FII a was determined by atomic absorption spectroscopy. Results: After incubation with FII a (0.25 g/L), Aα‐, Bβ‐ and γ‐chains of fibrinogen disappeared within 5 min, 30 min, and 8 h, respectively. The molecular weights of major degradation products were 45 000 and 41 000, which were different from those bands produced by plasmin. The fibrinogenolytic activity of FII a was strongly inhibited by ethylenediamine tetraacetic acid (EDTA), ethyleneglycol tetraacetic acid (EGTA), dithiothreitol and cysteine, but not by phenylmethyl‐sulfonyl fluoride and soybean trypsin inhibitor. Zinc (3171±25 mg/kg), potassium (489±17 mg/kg) and calcium (319±13 mg/kg) were found in FII a . Zn 2+ , Ca 2+ and Mg 2+ could recover the fibrinogenolytic activity of FII a , which was inhibited by EDTA. Only Ca 2+ could recover the fibrinogenolytic activity inhibited by EGTA. Conclusion: FII a can degrade the Aα‐, Bβ‐ and γ‐chains of fibrinogen. FII a is a metalloproteinase, and Zn 2+ , Ca 2+ , and disulfide bonds are necessary for its fibrinogenolytic activity.