Purification and characterization of a protein inhibitor of calcium-dependent proteases from rat liver.
Purification and characterization of a protein inhibitor of calcium-dependent proteases from rat liver.
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DOI：
10.1016/0003-9861(84)90592-7
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发表时间：
1984-08
影响因子：
3.9
通讯作者：
G. Demartino;D. E. Croall
中科院分区：
文献类型：
--
作者：
G. Demartino;D. E. Croall
大鼠肝脏的可溶性提取物含有钙依赖性蛋白酶的蛋白质抑制剂。该抑制剂的表观Mr= 250,000，并在EGTA 存在下通过凝胶过滤色谱法与钙依赖性蛋白酶分离。该抑制剂已使用与 Affi-Gel 15 共价连接的钙依赖性蛋白酶通过亲和层析进行纯化。该抑制剂以钙依赖性方式与该亲和树脂特异性结合，并在 EDTA 或 EGTA 存在的情况下洗脱。纯化的抑制剂在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳上显示为单一蛋白质，Mr=125,000。据推测它在非变性条件下是二聚体。该抑制剂可抑制大鼠肝脏以及其他组织和物种的两种钙依赖性蛋白酶中的每一种。然而，它对测试的任何其他蛋白酶没有影响。
Soluble extracts of rat liver contain a protein inhibitor of calcium-dependent proteases. The inhibitor has an apparentMr= 250,000 and is separated from the calcium-dependent proteases by gel-filtration chromatography in the presence of EGTA. The inhibitor has been purified by affinity chromatography using a calcium-dependent protease covalently linked to Affi-Gel 15. The inhibitor specifically binds to this affinity resin in a calcium-dependent manner and elutes in the presence of EDTA or EGTA. The purified inhibitor appears as a single protein withMr= 125,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Presumably it is a dimer under nondenaturing conditions. The inhibitor inhibits each of two calcium-dependent proteases from rat liver and from other tissues and species. However, it has no effect on any other protease tested.