Homogeneous purification of human recombinant GST-Akt/PKB from Sf9 cells
Homogeneous purification of human recombinant GST-Akt/PKB from Sf9 cells
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DOI：
10.1006/prep.1999.1102
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发表时间：
1999-10-01
影响因子：
1.6
通讯作者：
Ferrari, S
中科院分区：
文献类型：
--
作者：
Fabbro, D;Batt, D;Ferrari, S
调节细胞状态的过多的细胞外刺激导致参与相对较少的负责将信号转导到细胞内部的途径。一种这样的途径是磷脂酰肌醇3-激酶（PI-3 K）的活化，其导致膜限制性第二信使聚磷脂酰肌醇3 '-磷酸的产生。Akt/蛋白激酶B（PKB）是由3 '-磷酸化肌醇脂质激活的酶。本文描述了在Sf 9细胞中表达和一步纯化人重组GST-PKB的方案。该方案允许产生大量均匀纯化的具有高比活性的GST-PKB，用于高通量筛选或结构研究。(C)北京：科学出版社.
The plethora of extracellular stimuli modulating the status of a cell results in the engagement of relatively few pathways responsible for transducing signals to the interior of the cell. One such pathway is the activation of phosphatidylinositol 3-kinase (PI-3K), which results in the generation of a membrane-restricted second messenger, polyphosphatidylinositide 3'-phosphate. Among the enzymes activated by 3'-phosphorylated inositol lipids is Akt/protein kinase B (PKB), Here we describe a protocol for the expression and one-step purification of human recombinant GST-PKB in Sf9 cells. This scheme allows generating large amounts of homogeneously purified GST-PKB with high specific activity to be employed in high-throughput screening or structural studies. (C) 1999 Academic Press.