AI Article Synopsis
- The amber suppression technique was utilized to genetically incorporate the allysine precursor N -(4-azidobenzoxycarbonyl)-δ,ϵ-dehydrolysine in E. coli, enabling the creation of proteins with specific lysine dimethylation.
- The research led to the synthesis of modified proteins, such as dimethyl-histone H3 and p53, which were used to investigate the activities of epigenetic enzymes like LSD1 and Tip60.
- Results showed that LSD1 effectively demethylates certain dimethylated histones (H3K4me2 and H3K9me2) while being ineffective on others, and that methylation of p53 at K372 activates Tip60
Article Abstract
Using the amber suppression approach, N -(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.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5206893 | PMC |
| http://dx.doi.org/10.1002/anie.201609452 | DOI Listing |
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