脊椎动物胶原酶的分光光度测定法。

Spectrophotometric assay for vertebrate collagenase.

作者信息

Weingarten H, Feder J

出版信息

Anal Biochem. 1985 Jun;147(2):437-40. doi: 10.1016/0003-2697(85)90294-5.

Abstract

Collagenase from normal human skin fibroblasts was found to catalyze the hydrolysis of esters and thio esters. This observation led to the development of a rapid, sensitive, continuous spectrophotometric assay for vertebrate collagenase using the thio peptolide Ac-ProLeuGly-S-LeuLeuGly-OC2H5 as substrate in the presence of 4,4'-dithiodipyridine or Ellman's Reagent. A Km of 0.004 M and a kcat of 370,000 h-1 were determined for the thio peptolide-enzyme reaction. The method is able to detect collagenase at concentrations as low as 2 ng/ml.

摘要

人们发现，来自正常人皮肤成纤维细胞的胶原酶能够催化酯类和硫酯的水解反应。这一发现促使人们开发出一种快速、灵敏、连续的分光光度法，用于检测脊椎动物胶原酶，该方法使用硫肽化合物Ac-ProLeuGly-S-LeuLeuGly-OC2H5作为底物，并在4,4'-二硫代二吡啶或埃尔曼试剂存在的条件下进行。硫肽化合物与酶反应的米氏常数（Km）为0.004 M，催化常数（kcat）为370,000 h-1。该方法能够检测低至2 ng/ml浓度的胶原酶。