Recombinant production of fibrinogenase IV from Agkistrodon acutus venom and its preliminary evaluation.
A novel metalloproteinase, recombinant fibrinogenase IV (rFIVa), was expressed and purified from Agkistrodon acutus venom. It is a single-chain protein with an apparent molecular weight of 27 kDa. Western blot showed that it had a good immunological reaction against anti-FIVa rabbit serum. The kinetic parameters Km and Kcat of rFIVa on the substrate T6140 were 7.471 x 10(-4) mol/l and 5.103 x 10(-5) s(-1). RFIVa cleaved preferentially the alpha-chain, and the beta- and gamma-chains of fibrinogen were also cleaved when the incubation time was prolonged. The administration of rFIVa (1.8 and 5.4 mg/kg) to animals with acute blood-stasis model produced a decrease in fibrinogen to control values. To our knowledge, this is the first report of the expression, purification, and evaluation of recombinant fibrinogenase IV, which belongs to class P-I metalloproteinase from A. acutus venom.
Xu
Li-Peng
LP
Department of Pharmacology, Sun Yat-sen Medical School, Sun Yat-sen University, 74 Zhongshan II Road, Guangzhou, 510080, People's Republic of China.
Ma
Tao
T
Department of Pharmacology, Sun Yat-sen Medical School, Sun Yat-sen University, 74 Zhongshan II Road, Guangzhou, 510080, People's Republic of China.
Jiang
Wei-Jian
WJ
Department of Pharmacology, Sun Yat-sen Medical School, Sun Yat-sen University, 74 Zhongshan II Road, Guangzhou, 510080, People's Republic of China.
Qiu
Peng-Xin
PX
Department of Pharmacology, Sun Yat-sen Medical School, Sun Yat-sen University, 74 Zhongshan II Road, Guangzhou, 510080, People's Republic of China.
Wang
Xia
X
Department of Pharmacology, Sun Yat-sen Medical School, Sun Yat-sen University, 74 Zhongshan II Road, Guangzhou, 510080, People's Republic of China.
Su
Xing-Wen
XW
Department of Pharmacology, Sun Yat-sen Medical School, Sun Yat-sen University, 74 Zhongshan II Road, Guangzhou, 510080, People's Republic of China.
Yan
Guang-Mei
GM
Department of Pharmacology, Sun Yat-sen Medical School, Sun Yat-sen University, 74 Zhongshan II Road, Guangzhou, 510080, People's Republic of China. yanguangmei123@163.com.
D017836:Agkistrodon; D000818:Animals; D015153:Blotting, Western; D003001:Cloning, Molecular; D003435:Crotalid Venoms / Q000201:enzymology*; D005340:Fibrinogen / Q000378:metabolism; D007700:Kinetics; D023421:Models, Animal; D008970:Molecular Weight; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D012697:Serine Endopeptidases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D015003:Yeasts / Q000235:genetics / Q000378:metabolism
D003435:Crotalid Venoms; D011994:Recombinant Proteins; D005340:Fibrinogen; D012697:Serine Endopeptidases; C412879:venombin B