A small-molecule-protein interaction system with split-ubiquitin as sensor
A small-molecule-protein interaction system with split-ubiquitin as sensor
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DOI：
10.1002/cbic.200500544
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发表时间：
2006-06-01
期刊：
影响因子：
3.2
通讯作者：
Reichel, C
中科院分区：
文献类型：
--
作者：
Dirnberger, D;Unsin, G;Reichel, C
小分子受体的鉴定对于药物发现研究具有重要的药学意义。在后几个系统的识别蛋白质小分子相互作用已被开发。这些是经典酵母双杂交系统的修改，依赖于复合物核转位后的转录读出。在这里，我们提出了一种新的三杂交技术的基础上分裂泛素系统的分析蛋白质小分子相互作用的独立的核易位的复杂。该系统的性能进行了比较的方法的基础上的经典酵母双杂交系统，通过使用化学诱导剂的二聚化（CID）的氨甲蝶呤连接到地塞米松。通过改变化合物的接头长度以及通过比较融合蛋白的取向来解决空间问题。该系统进一步扩展到分析的人PCTAIRE蛋白激酶3，这是相关的细胞周期蛋白依赖性激酶（CDKs），一类重要的药物靶点的小分子抑制剂。
The identification of receptors for small molecules is of great pharmaceutical importance for drug-discovery research. Several systems for the identification of protein-small-molecule interactions have been developed in the post. These were modifications of the classical yeast two-hybrid system, relying on a transcriptional read-out following nuclear translocation of the complex. Here we present a novel three-hybrid technology based on the split-ubiquitin system for the analysis of protein-small-molecule interactions independently of a nuclear translocation of the complex. The performance of the system is compared to a method based on the classical yeast two-hybrid system by using a chemical inducer of dimerization (CID) comprised of methotrexate linked to dexamethasone. Steric issues are addressed by varying the linker length of the compounds, as well as by comparing the orientation of fusion proteins. The system is further extended to the analysis of a small-molecule inhibitor of human PCTAIRE protein kinase 3, which is related to cyclin-dependent kinases (CDKs), an important class of pharmaceutical targets.