Design, construction and characterization of prourokinase mutant engineered by introduction of special Lys-Gly-Asp-Trp-motif

DOI: 10.4236/abc.2013.32021   PDF   HTML   XML   4,417 Downloads   7,145 Views  

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.


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Jing, J. (2013) Design, construction and characterization of prourokinase mutant engineered by introduction of special Lys-Gly-Asp-Trp-motif. Advances in Biological Chemistry, 3, 164-169. doi: 10.4236/abc.2013.32021.

Conflicts of Interest

The authors declare no conflicts of interest.

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