Improved protein splicing using embedded split inteins

Protein Sci. 2018 Mar;27(3):614-619. doi: 10.1002/pro.3357. Epub 2018 Jan 17.

Abstract

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.

Keywords: chemical biology; intein splicing; protein engineering; protein semisynthesis.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Sequence
  • Inteins
  • Protein Engineering / methods*
  • Protein Splicing
  • Proteins / genetics
  • Proteins / metabolism*

Substances

  • Proteins