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• 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.