A promiscuous split intein with expanded protein engineering applications

Adam J. Stevens, Giridhar Sekar, Neel H. Shah, Anahita Z. Mostafavi, David Cowburn, Tom W. Muir

Research output: Contribution to journalArticlepeer-review

82 Scopus citations


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.

Original languageEnglish (US)
Pages (from-to)8538-8543
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Issue number32
StatePublished - Aug 8 2017

All Science Journal Classification (ASJC) codes

  • General


  • Chemical biology
  • Intein splicing
  • Protein engineering
  • Protein semisynthesis


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