A spectrophotometric assay for the determination of the catalytic efficiency of plasminogen activators using a slowly hydrolyzed plasmin substrate.
A spectrophotometric assay for the determination of the catalytic efficiency of plasminogen activators using a slowly hydrolyzed plasmin substrate.
复制标题
使用缓慢水解的纤溶酶底物测定纤溶酶原激活剂的催化效率的分光光度测定法。
DOI：
10.1006/abio.1995.1218
复制
发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Anderson,S
中科院分区：
文献类型：
--
作者：
Castro,MJ;Kingston,IB;Anderson,S
提出了一种测定纤溶酶原激活剂催化效率和活性的简便分光光度法。该检测系统包含激活剂、纤溶酶原和显色底物n -苯甲酰- l-精氨酸-对硝基苯胺（BAPA）。对于纤溶酶原，在非稳态的一级条件下，通过BAPA的水解来连续监测纤溶酶的产生。表观催化效率常数由表观一级活化速率常数计算得到。在稳态条件下，用本方法得到的结果与用不同体系测量kcat2和Km所测定的催化效率进行了比较。组织纤溶酶原激活剂和高分子量尿激酶作为模型激活剂。讨论了潜在的应用。
A simple spectrophotometric assay for the determination of the catalytic efficiency and activity of plasminogen activators is presented. The assay system contains activator, plasminogen, and the chromogenic substrate N-benzoyl-L-arginine-p-nitroanilide (BAPA). Plasmin production is monitored continuously by the hydrolysis of BAPA under non-steady-state, first-order conditions with respect to plasminogen. Apparent catalytic efficiency constants are calculated from the values obtained for the apparent first-order rate constant of activation. The results obtained with the present method were compared with the catalytic efficiency determined through the measurement of kcatand Km, using a different system, under steady-state conditions. Tissue plasminogen activator in the absence and presence of fibrinogen and high-molecular-weight urokinase were used as model activators. Potential applications are discussed.