A Versatile Approach for Site‐Specific Lysine Acylation in Proteins

Using amber suppression in coordination with a mutant pyrrolysyl‐tRNA synthetase‐tRNAPyl pair, azidonorleucine is genetically encoded in E. coli. Its genetic incorporation followed by traceless Staudinger ligation with a phosphinothioester allows the convenient synthesis of a protein with a site‐spe...

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Bibliographic Details
Published in:Angewandte Chemie 2017-02, Vol.129 (6), p.1665-1669
Main Authors: Wang, Zhipeng A., Kurra, Yadagiri, Wang, Xin, Zeng, Yu, Lee, Yan‐Jiun, Sharma, Vangmayee, Lin, Hening, Dai, Susie Y., Liu, Wenshe R.
Format: Article
Language:eng ; ger
Subjects:
Acetylation
Acylation
Amber
Amber-Unterdrückung
Azidonorleucin
Chemistry
E coli
Genetic code
Genetics
Government regulations
Histone H3
Histones
Lysinacylierung
Lysine
Post-translation
Protein biosynthesis
Proteinmodifikationen
Proteins
Staudinger-Ligation
Synthesis
Translation
tRNA
Ubiquitin
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Summary:Using amber suppression in coordination with a mutant pyrrolysyl‐tRNA synthetase‐tRNAPyl pair, azidonorleucine is genetically encoded in E. coli. Its genetic incorporation followed by traceless Staudinger ligation with a phosphinothioester allows the convenient synthesis of a protein with a site‐specifically installed lysine acylation. By simply changing the phosphinothioester identity, any lysine acylation type could be introduced. Using this approach, we demonstrated that both lysine acetylation and lysine succinylation can be installed selectively in ubiquitin and synthesized histone H3 with succinylation at its K4 position (H3K4su). Using an H3K4su‐H4 tetramer as a substrate, we further confirmed that Sirt5 is an active histone desuccinylase. Lysine succinylation is a recently identified post‐translational modification. The reported technique makes it possible to explicate regulatory functions of this modification in proteins. Azidonorleucin, eine Azid‐haltige Aminosäure, wurde genetisch codiert und in Modellproteine eingebaut. Dieser Einbau, zusammen mit einer nachfolgenden spurlosen Staudinger‐Ligation, erweitert erheblich die Möglichkeiten zur Synthese von Proteinen mit einer Vielzahl positionsspezifischer Lysinacylierungen.
ISSN:0044-8249
1521-3757
DOI:10.1002/ange.201611415