The inducible intein-mediated self-cleaving tag (Iist) system: A novel purification and amidation system for peptides and proteins

A. Sesilja Aranko, Hideo Iwaï*

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

Abstract

An efficient self-cleavable purification tag could be a powerful tool for purifying recombinant proteins and peptides without additional proteolytic processes using specific proteases. Thus, the intein-mediated self-cleavage tag was developed and has been commercially available as the IMPACT™ system. However, uncontrolled cleavages of the purification tag by the inteins in the IMPACT™ system have been reported, thereby reducing final yields. Therefore, controlling the protein-splicing activity of inteins has become critical. Here we utilized conditional protein splicing by salt conditions. We developed the inducible intein-mediated self-cleaving tag (IIST) system based on salt-inducible protein splicing of the MCM2 intein from the extremely halophilic archaeon, Halorhabdus utahensis and applied it to small peptides. Moreover, we described a method for the amidation using the same IIST system and demonstrated 15N-labeling of the C-terminal amide group of a single domain antibody (VHH).

Original languageEnglish
Article number5948
JournalMolecules
Volume26
Issue number19
DOIs
Publication statusPublished - 1 Oct 2021
MoE publication typeA1 Journal article-refereed

Keywords

  • C-terminal ami-dation
  • Conditional protein splicing
  • Halophilic intein
  • Isotopic labeling
  • NMR
  • Peptide purification
  • Protein splicing
  • Purification tag
  • Self-cleaving tag

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