A promiscuous split intein with expanded protein engineering applications.

Publication Year
2017

Type

Journal Article
Abstract

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.

Journal
Proc Natl Acad Sci U S A
Volume
114
Issue
32
Pages
8538-8543
Date Published
2017 Aug 08
ISSN Number
1091-6490
Alternate Journal
Proc Natl Acad Sci U S A
PMCID
PMC5559002
PMID
28739907