Alekseenko L P, Zolotov N N, Orekhovich V N
Vopr Med Khim. 1977 Mar-Apr;23(2):270-5.
A chromatographic method is developed for quantitative estimation of the collagenase-like enzyme (CLE) activity in extract of adenohypophysis and in preparations obtained during various steps of the enzyme isolation. The enzymatic hydrolysis of Cbz-Gly-Pro-Ala-Gly-Pro-Gly-OCH3 in presence of 2-mercaptoethanol at pH 8.0 was used as a pattern. The products formed were separated by chromatography on the paper; then they were stained with ninhydrin and converted into cupric complexes during extraction with ethanol; the optic density was measured at 510 nm. The optimal conditions for the enzymatic reaction were established. The method enabled to estimate the CLE activity in presence of prolyl carboxypeptidase. The specific effect of the CLE purified preparation on various synthetic peptides is discussed.
开发了一种色谱方法，用于定量测定腺垂体提取物以及酶分离各个步骤中获得的制剂中的胶原酶样酶（CLE）活性。以在pH 8.0的2-巯基乙醇存在下Cbz-Gly-Pro-Ala-Gly-Pro-Gly-OCH3的酶促水解作为模式。形成的产物通过纸色谱法分离；然后用茚三酮染色，并在用乙醇萃取过程中转化为铜络合物；在510nm处测量光密度。确定了酶促反应的最佳条件。该方法能够在脯氨酰羧肽酶存在下估计CLE活性。讨论了纯化的CLE制剂对各种合成肽的特异性作用。
