Measurement of the rate of plasmin action on synthetic substrates.
Measurement of the rate of plasmin action on synthetic substrates.
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测量纤溶酶对合成底物的作用速率。
DOI：
10.1016/s0021-9258(18)70394-3
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发表时间：
1958
期刊：
影响因子：
--
通讯作者：
P. S. Roberts
中科院分区：
文献类型：
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作者：
P. S. Roberts
Troll等人已经证明(1)纤溶酶(以非活性形式存在于血液中)可以水解酯的精氨酸和赖氨酸。作者用Form01滴定法跟踪纤溶酶对TAME(对甲苯磺酰-L-精氨酸甲酯)的作用，以及几种沉淀滴定方法(2)跟踪酶对L-赖氨酸甲酯的作用。他们还尝试了赫斯特林方法(3)，这是一种量热方法，基于酯类与碱性羟胺反应生成异羟肟酸的铁络合物。他们发现，这个程序给出了与滴定方法相同的结果，但他们没有给出实验细节，并表示滴定方法更可取，因为它可以在最佳底物浓度下使用。在这个实验室中，我们发现滴定方法，特别是LME的滴定方法，重复性差，主要是因为终点困难。我们修改了Hestrin方法，使TAME或LME的最佳底物浓度下的酶速率可以使用相同的标准试剂和程序来测定。通过这一过程，我们发现速率测量是可重复性的，平均偏差为速率的3%。
It has been shown by Troll et al.(1) that plasmin (present in the blood in an inactive form) can hydrolyze esters of arginine and lysine. These authors used a form01 titration method to follow the action of plasmin on TAME (p-toluenesulfonyl-L-arginine methyl ester) and several precipitation and titration methods (2) to follow the action of the enzyme on LME (L-lysine methyl ester). They also tried the Hestrin method (3), which is a calorimetric method based on the formation of a ferric complex of the hydroxamic acid resulting from the reaction of an ester with alkaline hydroxylamine. They found that this procedure gave the same results as the titration methods, but they gave no experimental details and stated that the titration method was preferable because it could be used at optimal substrate concentrations. In this laboratory, we have found that the titration methods, expecially for LME, give poor reproducibility, primarily because of difficult end points.We have modified the Hestrin method so that enzyme rates at optimal substrate concentrations of either TAME or LME can be determined by using the same standard reagents and procedures. With this procedure we find that the rate measurements are reproducible with an average deviation of 3 per cent of the rate.