Genetic Incorporation of Olefin Cross-Metathesis Reaction Tags for Protein Modification
Genetic Incorporation of Olefin Cross-Metathesis Reaction Tags for Protein Modification
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DOI：
10.1021/jacs.8b09433
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发表时间：
2018-11-07
影响因子：
15
通讯作者：
Davis, Benjamin G.
中科院分区：
文献类型：
--
作者：
Bhushan, Bhaskar;Lin, Yuya A.;Davis, Benjamin G.
烯烃交叉歧化反应(CM)是一种可行的烯烃类蛋白质修饰反应。虽然蛋白质中的烯丙基硫化物或硒侧链基序可以显著提高CM反应的速度，但到目前为止还没有有效的方法将它们选择性地遗传整合到蛋白质中。在这里，通过对含有非天然氨基酸的烯烃代谢结合的系统评价，我们发现S-烯丙基同型半胱氨酸是一种可遗传编码的蛋氨酸类似物，它不仅被翻译的细胞机加工，而且在蛋白质中也是特权的CM底物残基。通过这种方法，AHC被用于Met营养缺陷型大肠杆菌(B834(DE3))中有效的Met密码子重新分配以及人类细胞中蛋白质的代谢标记，并在几种代表性蛋白质中对CM起反应。这扩大了CM在蛋白质“标记和修改”功能化工具箱中的使用。
Olefin cross-metathesis (CM) is a viable reaction for the modification of alkene-containing proteins. Although allyl sulfide or selenide side-chain motifs in proteins can critically enhance the rate of CM reactions, no efficient method for their site-selective genetic incorporation into proteins has been reported to date. Here, through the systematic evaluation of olefin bearing unnatural amino acids for their metabolic incorporation, we have discovered S-allylhomocysteine (Ahc) as a genetically encodable Met analogue that is not only processed by translational cellular machinery but also a privileged CM substrate residue in proteins. In this way, Ahc was used for efficient Met codon reassignment in a Met-auxotrophic strain of E. coli (B834 (DE3)) as well as metabolic labeling of protein in human cells and was reactive toward CM in several representative proteins. This expands the use of CM in the toolkit for "tag-and-modify" functionalization of proteins.