Protein disulphide-isomerase from human placenta and rat liver. Purification and immunological characterization with monoclonal antibodies.
Protein disulphide-isomerase from human placenta and rat liver. Purification and immunological characterization with monoclonal antibodies.
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DOI：
10.1042/bj2410039
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发表时间：
1987
期刊：
影响因子：
--
通讯作者：
C. Kaetzel;C. K. Rao;M. Lamm
中科院分区：
文献类型：
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作者：
C. Kaetzel;C. K. Rao;M. Lamm
本文描述了人胎盘和大鼠肝蛋白二硫化物异构酶（PDI，EC 5.3.4.1）的纯化和一组抗这些蛋白的单克隆抗体的制备。人PDI和大鼠PDI的物理和酶性质相似;免疫学表征揭示了存在独特的，以及共享的抗原决定簇。虽然纯化的大鼠肝脏PDI存在三种形式略有不同的先生值，证据表明，多种形式代表一个单一的59,000-先生物种的蛋白水解降解产物。纯化的人PDI具有61，200的表观Mr。两种抗人PDI的单克隆抗体部分灭活了酶，其中一种在间接免疫沉淀中导致所有谷胱甘肽：胰岛素转氢酶活性从人胎盘的粗提取物中沉淀。HT-29人结肠癌细胞的免疫荧光实验结果与PDI在核膜和细胞质中的定位一致。
The purification of human placenta and rat liver protein disulphide-isomerase (PDI, EC 5.3.4.1) and the production of a panel of monoclonal antibodies against these proteins are described. The physical and enzymic properties of human PDI and rat PDI were similar; immunological characterization revealed the presence of unique, as well as shared, antigenic determinants. Although purified rat liver PDI was present as three forms differing slightly in Mr value, evidence was presented that the multiple forms represent proteolytic degradation products of a single 59,000-Mr species. Purified human PDI had an apparent Mr of 61,200. Two of the monoclonal antibodies against human PDI partially inactivated the enzyme, and one of these in indirect immunoprecipitation led to the precipitation of all glutathione:insulin transhydrogenase activity from a crude extract of human placenta. Results of immunofluorescence experiments with HT-29 human colon carcinoma cells were consistent with localization of PDI in the nuclear membrane and cell cytoplasm.