1种新型合成水蛭肽的纤溶动力学研究

发布时间：2018-07-28 21:08

【摘要】：目的研究水蛭肽Hirulog-s影响单链尿激酶型纤溶酶原激活剂和纤溶酶原相互活化作用考察新型合成水蛭肽的纤溶作用特性。方法构筑单链尿激酶型纤溶酶原激活剂(pro-uPA)和纤溶酶原(plg)相互活化反应体系,添加Hirulog-s成反应体系不同浓度,比较pro-uPA和plg相互活化反应体系中纤溶酶(plm)作用于pro-uPA反应的酶促反应动力学常数(Km,kcat),分析Hirulog-s对plm作用于pro-uPA反应的催化速率及亲和性。纤溶活性作用检测采用底物酶促反应法,酶促反应动力学常数的测定基于单链尿激酶型纤溶酶原激活剂激活纤溶酶原的反应。结果在plm作用于pro-uPA的反应体系中,30μmol·L~(-1) Hirulog-s与对照相比kcat值增大了3.1倍,Km值减小了1.6倍,表明Hirulog-s可以显著提高plm的最大催化速率同时增强亲和性。30μmol·L~(-1) Hirulog-s与对照相比kcat/Km值增大了4.75倍,表明Hirulog-s明显提高了plm的催化效率。通过活性对比表明Hirulog-s的纤溶活性强于水蛭肽Hirulog-1。结论 Hirulog-s促进单链尿激酶型纤溶酶原激活剂和纤溶酶原相互活化作用而发挥纤溶作用,新型合成水蛭肽从酶促反应动力学参数显示出优良的纤溶作用特性。
[Abstract]:Aim to investigate the effects of leech peptide Hirulog-s on the interaction between single chain urokinase plasminogen activator and plasminogen. Methods the system of single chain urokinase plasminogen activator (pro-uPA) and plasminogen (plg) interaction was constructed, and the different concentration of Hirulog-s was added to the system. The kinetic constants of pro-uPA reaction induced by plasminase (plm) (plm) were compared in the system of pro-uPA and plg interaction, and the catalytic rate and affinity of Hirulog-s to pro-uPA reaction by plm were analyzed. The activity of plasminogen was determined by substrate enzymatic reaction and the kinetic constant was determined based on the single chain urokinase type plasminogen activator activated plasminogen reaction. Results in the reaction system of pro-uPA treated with plm, the kcat value of 30 渭 mol L ~ (-1) Hirulog-s increased by 3.1-fold and decreased by 1.6-fold, which indicated that Hirulog-s could significantly increase the maximum catalytic rate of plm and enhance the affinity. 30 渭 mol L ~ (-1) Hirulog-s increased the kcat/Km value by 4.75 times compared with the control. The results show that Hirulog-s can obviously improve the catalytic efficiency of plm. The results showed that the fibrinolytic activity of Hirulog-s was stronger than that of Hirulog-1. Conclusion Hirulog-s promotes the interaction between single chain urokinase plasminogen activator and plasminogen and plays a role in fibrinolysis. The new synthesis of leech peptide shows excellent fibrinolytic effect from the kinetic parameters of enzymatic reaction.
【作者单位】：上海海洋大学食品科学与工程学院;赤峰学院附属医院;军事医学科学院生物工程研究所;
【基金】：国家自然科学基金项目(81341082,81502955)资助
【分类号】：R96

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