内源性钙蛋白酶抑制蛋白的分离

Isolation of Endogenous Calpastatin.

作者信息

De Tullio Roberta, Averna Monica

机构信息

Department of Experimental Medicine (DIMES)-Biochemistry Section, University of Genova, Genova, Italy.

Centre of Excellence for Biomedical Research (CEBR), University of Genova, Genova, Italy.

出版信息

Methods Mol Biol. 2019;1915:187-194. doi: 10.1007/978-1-4939-8988-1_14.

DOI:10.1007/978-1-4939-8988-1_14

PMID:30617804

Abstract

We here describe the purification of calpastatin from human erythrocytes. When calpastatin is purified from tissues, it is necessary to measure its inhibitory activity against calpain in the presence of Ca to specifically identify the protein. Thus, the purification steps necessary to obtain the inhibitor protein were originally designed to obtain calpain from the same tissue. For this reason, in addition to calpastatin purification, we also include a method for purifying human erythrocyte calpain and globin. We routinely use these two components for assaying calpastatin inhibition.

摘要

我们在此描述从人红细胞中纯化钙蛋白酶抑制蛋白的方法。当从组织中纯化钙蛋白酶抑制蛋白时，有必要在钙离子存在的情况下测量其对钙蛋白酶的抑制活性，以特异性鉴定该蛋白质。因此，最初设计用于获得抑制蛋白的纯化步骤是为了从同一组织中获得钙蛋白酶。出于这个原因，除了钙蛋白酶抑制蛋白的纯化外，我们还包括一种纯化人红细胞钙蛋白酶和珠蛋白的方法。我们通常使用这两种成分来测定钙蛋白酶抑制蛋白的抑制作用。