大肠杆菌表达的重组人PAI－1的纯化

纯化了工程细菌表达的可溶态和包含体形式的ｒｈＰＡＩ－１，纯度均达９８％，其ｒｈＰＡＩ－１蛋白质得率分别为１５％和１９％，比活性分别为３３５００ＩＵ／ｍｇ和２７７０００ＩＵ／ｍｇ。Ｎ端氨基酸序列分析显示，ｒｈＰＡＩ－１Ｎ端１５个氨基酸与天然ＰＡＩ－１完全一致；包含体复性研究表明，包含体的复性与复性蛋白的浓度及复性液中助溶剂的浓度密切相关。纯化的ｒｈＰＡＩ－１为分析ＰＡＩ－１结构与功能及探讨其临床应用提供了材料。

Purification of Recombinant Human PAI-1 Produced in E. coli

The inclusion body and soluble form of rhPAI-1 expressed in E. coli were purifiedwith a simple and effective protocol. The purity of PAI-1 was both up to 98%, the rhPAI-1 protein yielding was 19% and 15%, and specific activtirty of rhPAI-1 was 277 000 IU/mg and 33 500 IU/mg respectively. The sequence of N-terminal 15 amino acid residues of purified soluble rhPAI-1 was determined and found to be identical to the native protein. The renaturation of rhPAI-1 inclusion body is related to the protein and urea concentrations in the renaturation buffer. The purified rhPAI-1 may be applied to analysing its structure and function and to investigating its clinical effects.