Expanding the genetic code of Escherichia coli with phosphotyrosine

被引：67
作者：
Fan, Chenguang [1 ,2 ]
Ip, Kevan [2 ]
Soll, Dieter [2 ,3 ]
机构：
[1] Univ Arkansas, Dept Chem & Biochem, Fayetteville, AR 72701 USA
[2] Yale Univ, Dept Mol Biophys & Biochem, New Haven, CT 06511 USA
[3] Yale Univ, Dept Chem, New Haven, CT USA
来源：
FEBS LETTERS | 2016年 / 590卷 / 17期
关键词：
aminoacyl-tRNA synthetase; elongation factor; genetic code expansion; phosphatase; phosphotyrosine; tyrosine phosphorylation; AMINOACYL-TRANSFER-RNA; TYROSINE PHOSPHORYLATION; HUMAN GENOME; PHOSPHOSERINE; EVOLUTION; PROTEINS; ACID; EXPANSION; ORGANISM; AMBER;
D O I：
10.1002/1873-3468.12333
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
Protein phosphorylation is one of the most important post-translational modifications in nature. However, the site-specific incorporation of O-phosphotyrosine into proteins in vivo has not yet been reported. Endogenous phosphatases present in cells can dephosphorylate phosphotyrosine as a free amino acid or as a protein residue. Therefore, we deleted the genes of five phosphatases from the genome of Escherichia coli with the aim of stabilizing phosphotyrosine. Together with an engineered aminoacyl-tRNA synthetase (derived from Methanocaldococcus jannaschii tyrosyl-tRNA synthetase) and an elongation factor Tu variant, we were able to cotranslationally incorporate O-phosphotyrosine into the superfolder green fluorescent protein at a desired position in vivo. This system will facilitate future studies of tyrosine phosphorylation.
引用
收藏
页码：3040 / 3047
页数：8
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