Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 114, Issue 32, Pages 8538-8543Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1701083114
Keywords
chemical biology; protein engineering; intein splicing; protein semisynthesis
Categories
Funding
- National Institutes of Health [R37-GM086868, R01-GM107047, S10 OD016305-01A1]
- National Science Foundation [DGE-1148900, ACI-1053575]
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The protein trans-splicing (PTS) activity of naturally split inteins has found widespread use in chemical biology and biotechnology. However, currently used naturally split inteins suffer from an extein dependence, whereby residues surrounding the splice junction strongly affect splicing efficiency, limiting the general applicability of many PTS-based methods. To address this, we describe a mechanism-guided protein engineering approach that imbues ultrafast DnaE split inteins with minimal extein dependence. The resulting promiscuous inteins are shown to be superior reagents for protein cyclization and protein semisynthesis, with the latter illustrated through the modification of native cellular chromatin. The promiscuous inteins reported here thus improve the applicability of existing PTS methods and should enable future efforts to engineer promiscuity into other naturally split inteins.
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