Rat liver pyruvate carboxylase. Purification, detection and quantification of apo and holo forms by immuno-blotting and by an enzyme-linked immunosorbent assay.
Rat liver pyruvate carboxylase. Purification, detection and quantification of apo and holo forms by immuno-blotting and by an enzyme-linked immunosorbent assay.
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大鼠肝脏丙酮酸羧化酶。
DOI：
10.1042/bj2360527
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Mendez,A
中科院分区：
文献类型：
--
作者：
Ahmad,F;Ahmad,PM;Mendez,A
本文介绍了一种从大鼠肝线粒体中纯化丙酮酸羧化酶的简单方法。它是快速的，并提供了高纯度的丙酮酸羧化酶，具有优异的产率和再现性。最终的酶制剂似乎是均匀的，通过以下标准：洗脱行为的分子大小矩阵，SDS/聚丙烯酰胺凝胶电泳，Ouchterlony双扩散分析和蛋白质印迹。检测和定量纳克量的丙酮酸羧化酶（载脂蛋白和全息形式）在总组织匀浆免疫印迹和酶联免疫吸附测定。所提供的数据表明，在正常生理条件下（体内和体外），基本上所有的丙酮酸羧化酶分子都是生物素化的。
A simple scheme for the purification of pyruvate carboxylase from rat liver mitochondria is described. It is rapid and provides high-purity pyruvate carboxylase with excellent yield and reproducibility. The final enzyme preparations appear to be homogeneous by the following criteria: elution behaviour on molecular-sizing matrix, SDS/polyacrylamide-gel electrophoresis, Ouchterlony double-diffusion analysis and Western blotting. Detection and quantification of nanogram amounts of pyruvate carboxylase (apo and holo forms) in total tissue homogenates by immuno-blotting and by enzyme-linked immunosorbent assay are described. The data provided suggest that under normal physiological conditions (both in vivo and in vitro) essentially all the pyruvate carboxylase molecules are biotinylated.