A promiscuous split intein with expanded protein engineering applications.

TitleA promiscuous split intein with expanded protein engineering applications.
Publication TypeJournal Article
Year of Publication2017
AuthorsStevens, AJ, Sekar, G, Shah, NH, Mostafavi, AZ, Cowburn, D, Muir, TW
JournalProc Natl Acad Sci U S A
Volume114
Issue32
Pagination8538-8543
Date Published2017 08 08
ISSN1091-6490
KeywordsBacterial Proteins, Biotechnology, DNA Polymerase III, Exteins, Inteins, Models, Molecular, Nostoc, Protein Engineering, Protein Splicing, Synechocystis
Abstract

<p>The protein -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.</p>

DOI10.1073/pnas.1701083114
Alternate JournalProc. Natl. Acad. Sci. U.S.A.
PubMed ID28739907
PubMed Central IDPMC5559002
Grant ListR01 GM107047 / GM / NIGMS NIH HHS / United States
R37 GM086868 / GM / NIGMS NIH HHS / United States
S10 OD016305 / OD / NIH HHS / United States
T32 GM007388 / GM / NIGMS NIH HHS / United States