Novel biotin linker with alkyne and amino groups for chemical labelling of a target protein of a bioactive small molecule
Novel biotin linker with alkyne and amino groups for chemical labelling of a target protein of a bioactive small molecule
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DOI：
10.1016/j.bmcl.2017.12.055
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发表时间：
2018-02-15
影响因子：
2.7
通讯作者：
Takahashi, Kosaku
中科院分区：
文献类型：
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作者：
Anabuki, Tomoaki;Tsukahara, Miu;Takahashi, Kosaku
我们利用含有叠氮基团的小分子(叠氮探针)合成了一种新型的含有生物素、炔基和氨基的连接物(1)，用于鉴定靶蛋白。连接物中的乙炔通过叠氮-乙炔惠氏环加成反应与叠氮化物探针结合。蛋白质交联剂有效地将连接物和叠氮化物探针的结合物与目标蛋白质结合。Western blotting检测到共价结合的复合体。将连接物1应用于使用脱落酸受体RCAR/PYR/PYL(PYL)的模型系统。Western blotting成功地显示了接头1、叠氮化物探针和目的蛋白的交联物。这种识别目标蛋白的方法比以前开发的方法更有效，该方法使用第二个连接物与生物素、炔和二苯甲酮(连接物2)作用于光交联靶蛋白。本研究开发的系统是一种识别生物活性小分子靶蛋白的方法，不同于光亲和标记法。(C)2017爱思唯尔有限公司。保留所有权利。
We synthesized a novel linker (1) with biotin, alkyne and amino groups for the identification of target proteins using a small molecule that contains an azide group (azide probe). The alkyne in the linker bound the azide probe via an azide-alkyne Huisgen cycloaddition. A protein cross-linker effectively bound the conjugate of the linker and an azide probe with a target protein. The covalently bound complex was detected by western blotting. Linker 1 was applied to a model system using an abscisic acid receptor, RCAR/PYR/PYL (PYL). Cross-linked complexes of linker 1, the azide probes and the target proteins were successfully visualized by western blotting. This method of target protein identification was more effective than a previously developed method that uses a second linker with biotin, alkyne, and benzophenone (linker 2) that acts to photo-crosslink target proteins. The system developed in this study is a method for identifying the target proteins of small bioactive molecules and is different from photo-affinity labelling. (C) 2017 Elsevier Ltd. All rights reserved.