title: Expression, purification and characterization of recombinant plasminogen activator from Gloydius brevicaudus venom in Escherichia coli.
abstract: The plasminogen activator (PA) in snake venom, a serine protease, can convert plasminogen to active plasmin, indirectly causing the degradation of fibrin. It is difficult to purify sufficient snake venom PA (SV-PA) for clinical applications due to the low SV-PA content in venom. The gene encoding PA was obtained from the venom gland of Gloydius brevicaudus using RT-PCR with primers designed according to the N-terminal amino acids of G. brevicaudus venom PA (GBV-PA), was cloned into the prokaryotic expression vector pET-42a, and recombinant GBV-PA (rGBV-PA) was expressed via Isopropyl-β-d-1-Thiogalactopyranoside (IPTG) induction. Like human tissue PA, the purified renatured rGBV-PA could significantly reduce the rabbit plasma euglobulin lysis time (ELT) and prevent thrombus formation in the inferior vena cava of rats. Within the dosage range, the dosage and effects were positively correlated.
authors.affiliation: School of Pharmacy, Fujian Medical University, Fuzhou 350004, PR China; Department of Pharmacy, Fujian Medical University Union Hospital, Fuzhou 350001, PR China.
authors.forename: Jinhua
authors.initials: J
authors.lastname: Zhang
authors.forename: Wenli
authors.initials: W
authors.lastname: Meng
authors.forename: Chunhua
authors.initials: C
authors.lastname: Wang
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Wu
authors.forename: Guotu
authors.initials: G
authors.lastname: Wu
authors.forename: Yunlu
authors.initials: Y
authors.lastname: Xu
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D003435:Crotalid Venoms / Q000737:chemistry*; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005260:Female; D063285:Fibrin Clot Lysis Time; D005342:Fibrinolysis / Q000187:drug effects; D008297:Male; D010960:Plasminogen Activators / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D011817:Rabbits; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D020246:Venous Thrombosis / Q000473:pathology; D017819:Viperidae / Q000235:genetics*
chemical_list: D003435:Crotalid Venoms; D011994:Recombinant Proteins; D010960:Plasminogen Activators
keywords: Escherichia coli; Gloydius brevicaudus; Molecular clone; Plasminogen activator; Thrombosis