Genetically encoding an aliphatic diazirine for protein photocrosslinking
Genetically encoding an aliphatic diazirine for protein photocrosslinking
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DOI：
10.1039/c0sc00373e
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发表时间：
2011-01-01
期刊：
影响因子：
8.4
通讯作者：
Deiters, Alexander
中科院分区：
文献类型：
--
作者：
Chou, Chungjung;Uprety, Rajendra;Deiters, Alexander
光交联是一种重要的方法，可以发现和详细研究蛋白质-蛋白质、蛋白质-寡核苷酸和蛋白质-小分子相互作用，具有高时间和空间分辨率。该方法普遍应用的一个主要限制是将高效的脂肪族光交联探针引入感兴趣的蛋白质中。在这里，我们报告了一种新的脂肪族重氮嘧啶氨基酸及其基因编码，位点特异性结合到细菌和哺乳动物细胞的蛋白质中。此外，我们在体外和体内证明了一种测试蛋白的有效光交联。
Photocrosslinking is an important approach that allows discovery and detailed investigation of protein-protein, protein-oligonucleotide, and protein-small molecule interactions with high temporal and spatial resolution. A major limitation to the universal application of this methodology is the site-specific introduction of efficient aliphatic photocrosslinking probes into proteins of interest. Here, we report a novel aliphatic diazirine amino acid and its genetically encoded, site-specific incorporation into proteins in bacterial and mammalian cells. Furthermore, we demonstrate efficient photocrosslinking of a test protein in vitro and in vivo.