Improving the Solubility of Artificial Ligands of Streptavidin to Enable More Practical Reversible Switching of Protein Localization in Cells
Improving the Solubility of Artificial Ligands of Streptavidin to Enable More Practical Reversible Switching of Protein Localization in Cells
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DOI：
10.1002/cbic.201600640
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发表时间：
2017-02
期刊：
影响因子：
3.2
通讯作者：
Ryo Tachibana;Takuya Terai;Gaelle Boncompain;S. Sugiyama;Nae Saito;F. Perez;Y. Urano
中科院分区：
文献类型：
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作者：
Ryo Tachibana;Takuya Terai;Gaelle Boncompain;S. Sugiyama;Nae Saito;F. Perez;Y. Urano
化学诱导剂可以控制活细胞中靶蛋白的定位，是研究动态生物系统的有力工具。我们最近报道了使用选择性钩或“RUSH”系统通过添加/洗掉链霉亲和素（ALiS）的小分子人工配体来保留感兴趣的蛋白质的可逆定位变化。然而，先前开发的ali的效用受到水溶解度的限制。在这里，我们克服了这个问题，通过X射线晶体结构引导设计了一种更可溶性的ALiS衍生物（ALiS‐3），它保留了足够的链亲和素结合亲和力，用于RUSH系统。详细表征了ALiS‐3 -链亲和素的相互作用。ALiS‐3是动态控制活细胞内质网和高尔基体之间α‐甘露糖苷酶II定位的方便有效的工具。
Chemical inducers that can control target‐protein localization in living cells are powerful tools to investigate dynamic biological systems. We recently reported the retention using selective hook or “RUSH” system for reversible localization change of proteins of interest by addition/washout of small‐molecule artificial ligands of streptavidin (ALiS). However, the utility of previously developed ALiS was restricted by limited solubility in water. Here, we overcame this problem by X‐ray crystal structure‐guided design of a more soluble ALiS derivative (ALiS‐3), which retains sufficient streptavidin‐binding affinity for use in the RUSH system. The ALiS‐3–streptavidin interaction was characterized in detail. ALiS‐3 is a convenient and effective tool for dynamic control of α‐mannosidase II localization between ER and Golgi in living cells.