北京大学陈鹏团队报道了基因编码的生物正交色氨酸在活细胞中衰减。相关研究成果于2024年2月28日发表在《自然—化学》。

色氨酸（Trp）通过其π系统和吲哚N–H基团在调节蛋白质结构、相互作用和功能方面发挥着关键作用。阻断和恢复Trp相互作用的通用方法将使各种含Trp的蛋白质能够在体内获得功能操作，但产生这样的平台仍然具有挑战性。

该文中，研究人员开发了一种基因编码的N1乙烯基笼状Trp，它能够通过优化的反电子需求Diels–Alder反应快速和生物正交衰变，实现活细胞中感兴趣的蛋白质上的Trp位点特异性激活。这种基因编码的笼状色氨酸（Trp-CAGE）策略的化学激活能够精确激活潜在的各种重要分子相互作用的感兴趣的Trp。

研究证明了Trp-CAGE在各种蛋白质家族中的效用，如过氧化氢酶过氧化物酶和激酶，作为翻译起始物和翻译后修饰读取器，能够以时间控制的方式调节表观遗传信号。结合计算机辅助预测，该策略为28000多种候选蛋白质在其天然细胞环境中的生物正交Trp激活铺平了道路。

附：英文原文

Title: Genetically encoded bioorthogonal tryptophan decaging in living cells

Author: Zhu, Yuchao, Ding, Wenlong, Chen, Yulin, Shan, Ye, Liu, Chao, Fan, Xinyuan, Lin, Shixian, Chen, Peng R.

Issue&Volume: 2024-02-28

Abstract: Tryptophan (Trp) plays a critical role in the regulation of protein structure, interactions and functions through its π system and indole N–H group. A generalizable method for blocking and rescuing Trp interactions would enable the gain-of-function manipulation of various Trp-containing proteins in vivo, but generating such a platform remains challenging. Here we develop a genetically encoded N1-vinyl-caged Trp capable of rapid and bioorthogonal decaging through an optimized inverse electron-demand Diels–Alder reaction, allowing site-specific activation of Trp on a protein of interest in living cells. This chemical activation of a genetically encoded caged-tryptophan (Trp-CAGE) strategy enables precise activation of the Trp of interest underlying diverse important molecular interactions. We demonstrate the utility of Trp-CAGE across various protein families, such as catalase-peroxidases and kinases, as translation initiators and posttranslational modification readers, allowing the modulation of epigenetic signalling in a temporally controlled manner. Coupled with computer-aided prediction, our strategy paves the way for bioorthogonal Trp activation on more than 28,000 candidate proteins within their native cellular settings.

DOI: 10.1038/s41557-024-01463-7

Source: https://www.nature.com/articles/s41557-024-01463-7