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A Genetically Encoded Allysine for the Synthesis of Proteins with Site‐Specific Lysine Dimethylation
Author(s) -
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.
Publication year - 2017
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201609452
Subject(s) - acetylation , histone h3 , demethylase , lysine , acetyltransferase , histone , histone acetyltransferase , methylation , chemistry , biochemistry , epigenetics , methyltransferase , histone methyltransferase , microbiology and biotechnology , biology , dna , amino acid , gene
Using the amber suppression approach, N ϵ ‐(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.