利用扩展遗传密码实现磷酸酪氨酸的位点特异性掺入。
Site-specific incorporation of phosphotyrosine using an expanded genetic code.
作者信息
Hoppmann Christian, Wong Allison, Yang Bing, Li Shuwei, Hunter Tony, Shokat Kevan M, Wang Lei
机构信息
Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, USA.
Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, California, USA.
出版信息
Nat Chem Biol. 2017 Aug;13(8):842-844. doi: 10.1038/nchembio.2406. Epub 2017 Jun 12.
Access to phosphoproteins with stoichiometric and site-specific phosphorylation status is key to understanding the role of protein phosphorylation. Here we report an efficient method to generate pure, active phosphotyrosine-containing proteins by genetically encoding a stable phosphotyrosine analog that is convertible to native phosphotyrosine. We demonstrate its general compatibility with proteins of various sizes, phosphotyrosine sites and functions, and reveal a possible role of tyrosine phosphorylation in negative regulation of ubiquitination.
获得具有化学计量和位点特异性磷酸化状态的磷蛋白是理解蛋白质磷酸化作用的关键。在此，我们报告了一种通过基因编码一种可转化为天然磷酸酪氨酸的稳定磷酸酪氨酸类似物来生成纯的、有活性的含磷酸酪氨酸蛋白的有效方法。我们证明了它与各种大小、磷酸酪氨酸位点和功能的蛋白质具有普遍兼容性，并揭示了酪氨酸磷酸化在泛素化负调控中的可能作用。
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