期刊
SCIENCE
卷 354, 期 6312, 页码 623-626出版社
AMER ASSOC ADVANCEMENT SCIENCE
DOI: 10.1126/science.aah4428
关键词
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资金
- National Research Foundation of Korea (KAIST Systems Healthcare) [2011-0020322, 2014M3A6A4075060, 2012M3A9B4027956, 2012M3A9C6049937]
- U.S. National Institute for General Medical Sciences [GM22854-42]
- National Research Foundation of Korea [2014M3A6A4075060, 2012M3A9B4027956, 2012M3A9C6049937] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
Many essential biological processes are controlled by posttranslational protein modifications. The inability to synthetically attain the diversity enabled by these modifications limits functional studies of many proteins. We designed a three-step approach for installing authentic posttranslational modifications in recombinant proteins. We first use the established O-phosphoserine (Sep) orthogonal translation system to create a Sep-containing recombinant protein. The Sep residue is then dephosphorylated to dehydroalanine (Dha). Last, conjugate addition of alkyl iodides to Dha, promoted by zinc and copper, enables chemoselective carbon-carbon bond formation. To validate our approach, we produced histone H3, ubiquitin, and green fluorescent protein variants with site-specific modifications, including different methylations of H3K79. The methylated histones stimulate transcription through histone acetylation. This approach offers a powerful tool to engineer diverse designer proteins.
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