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Simulation, construction and characterization of a multi-functional thrombolytic agent with anti-thrombosis activity
Authors:Weiran Yu and Jian Jing
Institution:(1) Academy of Military Medicine, Beijing Institute of Bioengineering, Beijing, 100071, China;(2) Department of Biochemistry and Molecular Biology, Beijing Key Laboratory, Beijing Normal University, Beijing, 100875, China
Abstract:Prourokinase (scu-PA), a thrombolytic agent, was inserted between Gly118 and Ile119 with foreign antithrombosis functional motif (Lys-Gly-Asp-Trp-motif) to construct a multi-functional chimeric molecule. The molecular model of a chimera was simulated and predicted. The recombinant chimeric protein was expressed by the baculovirus-insect cell expression system and purified by affinity chromatography. The physico-chemical characteristics of the chimeric molecule were assayed. The thrombolytic activity was determined to be 90000 IU/mg of fibrinolytic special activity by the fibrin-plate method. The anti-thrombosis activities were also assayed with IC50 of 9.6 μM by an inhibition test of ADP-induced platelet aggregation.
Keywords:anti-thrombosis  thrombolytic agent  insect cell  baculovirus  prourokinase (scu-PA)  foreign functional motif
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