Substrate specificities of inteins investigated by QuickDrop-cassette mutagenesis

Jesper S. Oeemig, Hannes M. Beyer, A. Sesilja Aranko, Justus Mutanen, Hideo Iwaï*

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

5 Citations (Scopus)
52 Downloads (Pure)


Inteins catalyze self-excision from host precursor proteins while concomitantly ligating the flanking substrates (exteins) with a peptide bond. Noncatalytic extein residues near the splice junctions, such as the residues at the −1 and +2 positions, often strongly influence the protein-splicing efficiency. The substrate specificities of inteins have not been studied for many inteins. We developed a convenient mutagenesis platform termed “QuickDrop”-cassette mutagenesis for investigating the influences of 20 amino acid types at the −1 and +2 positions of different inteins. We elucidated 17 different profiles of the 20 amino acid dependencies across different inteins. The substrate specificities will accelerate our understanding of the structure–function relationship at the splicing junctions for broader applications of inteins in biotechnology and molecular biosciences.

Original languageEnglish
Pages (from-to)3338-3355
Number of pages18
JournalFEBS Letters
Issue number20
Early online date17 Aug 2020
Publication statusPublished - 1 Oct 2020
MoE publication typeA1 Journal article-refereed


  • intein
  • mutagenesis
  • protein ligation
  • protein splicing
  • substrate specificity


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