A Genetically Encoded Allysine for the Synthesis of Proteins with Site‐Specific Lysine Dimethylation

Using the amber suppression approach, Nϵ‐(4‐azidobenzoxycarbonyl)‐δ,ϵ‐dehydrolysine, an allysine precursor is genetically encoded in E. coli. Its genetic incorporation followed by two sequential biocompatible reactions allows convenient synthesis of proteins with site‐specific lysine dimethylation....

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Bibliographic Details
Published in:Angewandte Chemie International Edition 2017-01, Vol.56 (1), p.212-216
Main Authors: Wang, Zhipeng A., Zeng, Yu, Kurra, Yadagiri, Wang, Xin, Tharp, Jeffery M., Vatansever, Erol C., Hsu, Willie W., Dai, Susie, Fang, Xinqiang, Liu, Wenshe R.
Format: Article
Language:English
Subjects:
2-Aminoadipic Acid - analogs & derivatives
2-Aminoadipic Acid - genetics
Acetylation
allysine
amber suppression
Biocompatibility
Chemical synthesis
dimethyllysine
E coli
Escherichia coli - genetics
Genetic Code
genetic code expansion
Histone acetyltransferase
Histone H3
Histones - chemistry
Histones - genetics
Humans
Lysine
Lysine - analogs & derivatives
Lysine - chemistry
Lysine - genetics
lysine dimethylation
Methylation
Models, Molecular
Mutagenesis, Site-Directed - methods
p53 Protein
Protein Processing, Post-Translational
Proteins
Tumor Suppressor Protein p53 - chemistry
Tumor Suppressor Protein p53 - genetics
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Summary:Using the amber suppression approach, Nϵ‐(4‐azidobenzoxycarbonyl)‐δ,ϵ‐dehydrolysine, an allysine precursor is genetically encoded in E. coli. Its genetic incorporation followed by two sequential biocompatible reactions allows convenient synthesis of proteins with site‐specific lysine dimethylation. Using this approach, dimethyl‐histone H3 and p53 proteins have been synthesized and used to probe functions of epigenetic enzymes including histone demethylase LSD1 and histone acetyltransferase Tip60. We confirmed that LSD1 is catalytically active toward H3K4me2 and H3K9me2 but inert toward H3K36me2, and methylation at p53 K372 directly activates Tip60 for its catalyzed acetylation at p53 K120. Expedient protein synthesis: An allysine precursor, Nϵ‐(4‐azidobenzoxycarbonyl)‐δ,ϵ‐dehydrolysine is genetically encoded in E. coli. Its incorporation followed by Staudinger reduction and reductive amination allows the synthesis of proteins with site‐specific lysine dimethylation.
ISSN:1433-7851
1521-3773
DOI:10.1002/anie.201609452