%0 Journal Article
%T A prourokinase-RGDS chimera——Construction, expression and characterization
%A 钱斌
%A 孙迎庆
%A 郭雁
%A 党昕
%A 茹炳根
%J 中国科学C辑(英文版)
%D 1999
%I Springer
%X A tetrapeptide, RGDS, was inserted into proUK kringle domain G118-L119 by the construction of a mutant proUK-RGDS gene. The gene was expressed in the baculovirus expression system. Immunoaffinity chromatography was used to purify the chimera and protein with purity over 90% was achieved. The chimera was tested for its platelet membrane binding function and showed a calcium-dependent platelet binding activity. Amidolytic activity of the chimera was tested. The result indicated that specific amidolytic activity of plasmin activated chimera was 62000 IU/mg, comparable to the previously reported 65 355 IU/mg of plasmin activated natural proUK. Activation of plasminogen by the chimera after plasmin treatment followed Micbieal-Menten kinetics, and the Km was 0.97 μmol/L, which was also comparable to 1.64 μmol/L of native urokinase. The chimera also showed intensive ability to inhibit platelet aggregation in vitro. These results indicate that this chimera might be useful as a bifunctional thrombolytic agent.
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=180CF3A72E750F3261A8A60EDC957784&aid=642994E0336B04261A197176704D92B0&yid=B914830F5B1D1078&vid=ECE8E54D6034F642&iid=38B194292C032A66&sid=0F7768518993EDDE&eid=4FE459D71E3BF8EB&journal_id=1674-7305&journal_name=ScienceChina.Lifesciences&referenced_num=0&reference_num=0