Addition of the keto functional group to the genetic code of Escherichia coli
Addition of the keto functional group to the genetic code of Escherichia coli
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DOI：
10.1073/pnas.0234824100
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发表时间：
2003-01-07
影响因子：
11.1
通讯作者：
Schultz, PG
中科院分区：
文献类型：
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作者：
Wang, L;Zhang, ZW;Schultz, PG
尽管酮基是有机化学中最通用的官能团，但它在基因编码的氨基酸中不存在。为了克服蛋白质生物合成的这种天然限制，我们进化出了一种正交 tRNA 合成酶对，可以将酮基氨基酸（对乙酰基-L-苯丙氨酸）有效掺入大肠杆菌中的蛋白质中，并响应琥珀无义密码子，并具有高翻译保真度。为了证明这种酮氨基酸的实用性，我们用它用小分子荧光团和生物素衍生物选择性地修饰蛋白质。这种额外的基因编码氨基酸应该会极大地扩展我们在体外和活细胞中操纵蛋白质结构和功能的能力。
Although the keto group is the most versatile of the functional groups in organic chemistry, it is absent in the genetically encoded amino acids. To overcome this natural limitation on protein biosynthesis, we have evolved an orthogonal tRNA-synthetase pair that makes possible the efficient incorporation of a keto amino acid, p-acetyl-L-phenylalanine, into proteins in E. coli with high translational fidelity in response to the amber nonsense codon. To demonstrate the utility of this keto amino acid, we have used it to modify a protein selectively with a small molecule fluorophore and biotin derivative. This additional genetically encoded amino acid should greatly expand our ability to manipulate protein structure and function both in vitro and in living cells.