用于SH2结构域结合分析的可溶性STAT5b/STAT3蛋白的表达与纯化
Expression and Purification of Soluble STAT5b/STAT3 Proteins for SH2 Domain Binding Assay.
作者信息
Asai Akira, Takakuma Kazuyuki
机构信息
Center for Drug Discovery, Graduate School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka, 422-8526, Japan.
Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, Saitama, Japan.
出版信息
Methods Mol Biol. 2017;1555:163-172. doi: 10.1007/978-1-4939-6762-9_9.
When a large hydrophobic full-length protein is expressed in bacteria, it is often challenging to obtain recombinant proteins in the soluble fraction. One way to overcome this challenge is expression of deletion mutants that have improved solubility while maintaining biological activity. In this chapter, we describe a protocol for expression of truncated forms of STAT5b and STAT3 proteins that are soluble and retain SH2-mediated activity for phospho-Tyr peptide recognition.
当一个大的疏水性全长蛋白在细菌中表达时，要在可溶部分获得重组蛋白通常具有挑战性。克服这一挑战的一种方法是表达缺失突变体，这些突变体在保持生物活性的同时具有改善的溶解性。在本章中，我们描述了一种表达STAT5b和STAT3蛋白截短形式的方案，这些截短形式是可溶的，并保留了对磷酸化酪氨酸肽识别的SH2介导的活性。
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