Title | A promiscuous split intein with expanded protein engineering applications. |
Publication Type | Journal Article |
Year of Publication | 2017 |
Authors | Stevens, AJ, Sekar, G, Shah, NH, Mostafavi, AZ, Cowburn, D, Muir, TW |
Journal | Proc Natl Acad Sci U S A |
Volume | 114 |
Issue | 32 |
Pagination | 8538-8543 |
Date Published | 2017 Aug 08 |
ISSN | 1091-6490 |
Keywords | Bacterial 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> |
DOI | 10.1073/pnas.1701083114 |
Alternate Journal | Proc Natl Acad Sci U S A |
PubMed ID | 28739907 |
PubMed Central ID | PMC5559002 |
Grant List | R01 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 |