Open AccessPurification and partial characterizations of coagulant protein Fla from Daboia russelli siamensis (Myanmar) venom 1
Author(s) -
SUN Huanhuan,
CHEN Qi,
LIN Xi,
CHEN Jiashu,
QIU Pengxin,
YAN Guangmei
Publication year - 2007
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.1745-7254.2007.00616.x
Subject(s) - venom , sephadex , size exclusion chromatography , chromatography , thrombin , partial thromboplastin time , chemistry , factor x , fibrinogen , isoelectric point , coagulation , biochemistry , biology , enzyme , medicine , immunology , platelet , psychiatry
Aim: To purify and characterize the coagulant protein FIa from Daboia russelli siamensis (Myanmar) venom. Methods: FIa was purified from Daboia russelli siamensis (Myanmar) venom by ion‐exchange chromatography on CM‐Sephadex C‐50, and gel filtration on Sephadex G‐75 and a Superdex 75 column. The hemostatic activity of FIa was determined by the method of Williams and Esnouf. The specific chromogenic substrates were used respectively to determine the activation of factor X and prothrombin. The fibrinogen‐clotting activity of FIa was determined by the method of Gao et al. Normal saline was used as a negative control while factor Xa and thrombin were used as positive controls, respectively. Results: FIa, a coagulant protein, was achieved by ion‐exchange chromatography and gel filtration with a molecular weight of 34 479 and an isoelectric point of 7.2. FIa was shown to have strong hemostatic activity. The hemostatic activity of 0.5 mg FIa was equal to that of 1.5625 u thrombin. FIa primarily activated factor X, however, had no influence on prothrombin, nor did it cleave or clot fibrinogen. Conclusion: FIa is a factor X‐activating enzyme, which could activate factor X to factor Xa, but has no effect on prothrombin and fibrinogen.