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猪胰蛋白酶原(羧肽酶A)两种形式及其单体羧肽酶A的制备性分离

Preparative isolation of the two forms of pig pancreatic pro-(carboxypeptidase A) and their monomeric carboxypeptidases A.

作者信息

Vilanova M, Vendrell J, López M T, Cuchillo C M, Avilés F X

出版信息

Biochem J. 1985 Aug 1;229(3):605-9. doi: 10.1042/bj2290605.

Abstract

A method is reported for the preparative isolation of the two forms of pro-(carboxypeptidase A) from pig pancreas: the monomer and the binary complex with pro-(proteinase E). This method, which is mainly based on chromatography on DEAE-Sepharose at pH 5.7, allows these proenzymes to be prepared more quickly and in safer conditions than with other reported methods. Undegraded and homogeneous carboxypeptidase A1 and A2 species (peptidyl-L-amino acid hydrolase, EC 3.4.17.1), in monomeric forms with high specific activity, are also obtained in high yield by controlled trypsin activation of either of the pro-(carboxypeptidases A) followed by chromatography on DEAE-Sepharose at pH 5.8 under dissociating conditions (7 M-urea).

摘要

本文报道了一种从猪胰腺中制备分离两种形式的前羧肽酶A的方法:单体形式以及与前蛋白酶E的二元复合物形式。该方法主要基于在pH 5.7条件下的DEAE-琼脂糖柱层析,相较于其他已报道的方法,能更快速且在更安全的条件下制备这些酶原。通过对任一前羧肽酶A进行可控的胰蛋白酶激活,然后在解离条件(7M尿素)下于pH 5.8进行DEAE-琼脂糖柱层析,还能以高产率获得未降解且均一的羧肽酶A1和A2单体形式(肽基-L-氨基酸水解酶,EC 3.4.17.1),其具有高比活性。

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