TITLE:
Design, construction and characterization of prourokinase mutant engineered by introduction of special Lys-Gly-Asp-Trp-motif
AUTHORS:
Jian Jing
KEYWORDS:
Prourokinase; Lys-Gly-Asp-Trp-motif; Baculovirus-Insect Cell Expression System; Fibrinolytic Activity; Anti-Thrombosis Activity
JOURNAL NAME:
Advances in Biological Chemistry,
Vol.3 No.2,
April
30,
2013
ABSTRACT:
A recombinant prourokinase
chimera was constructed by introduction of Lys-Gly-Asp-Trp-motif between
Gly118 and Ile119 among the kringle domain. The structure of designed protein
was predicted and simulated. The recombinant prourokinase chimera was produced
in insect cell sf9 with baculovirus-expression vector and existed as active
form. Chimera protein was purified by affinity chromatography coupled with
antibody. The special activity of the chimera was 90,000 IU/mg detected by
fibrin plate determination. It was also shown that chimera inhibited
ADP-induced platelet aggregation in a concentration depenent manner. These
results showed the prourokinase chimera exhibited not only high fibrinolytic
activity but also had anti-thrombosis function.