Improved protein splicing using embedded split inteins

Josef A. Gramespacher, Adam J. Stevens, Robert E. Thompson, Tom W. Muir

Research output: Contribution to journalArticlepeer-review

15 Scopus citations


Naturally split inteins mediate a traceless protein ligation process known as protein trans-splicing (PTS). Although frequently used in protein engineering applications, the efficiency of PTS can be reduced by the tendency of some split intein fusion constructs to aggregate; a consequence of the fragmented nature of the split intein itself or the polypeptide to which it is fused (the extein). Here, we report a strategy to help address this liability. This involves embedding the split intein within a protein sequence designed to stabilize either the intein fragment itself or the appended extein. We expect this approach to increase the scope of PTS-based protein engineering efforts.

Original languageEnglish (US)
Pages (from-to)614-619
Number of pages6
JournalProtein Science
Issue number3
StatePublished - Mar 2018

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Biochemistry


  • chemical biology
  • intein splicing
  • protein engineering
  • protein semisynthesis


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