The Inducible Intein-Mediated Self-Cleaving Tag (IIST) System : A Novel Purification and Amidation System for Peptides and Proteins

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Aranko , A S & Iwaï , H 2021 , ' The Inducible Intein-Mediated Self-Cleaving Tag (IIST) System : A Novel Purification and Amidation System for Peptides and Proteins ' , Molecules , vol. 26 , no. 19 , 5948 . https://doi.org/10.3390/molecules26195948

Title: The Inducible Intein-Mediated Self-Cleaving Tag (IIST) System : A Novel Purification and Amidation System for Peptides and Proteins
Author: Aranko, A. Sesilja; Iwaï, Hideo
Contributor: University of Helsinki, Hideo Iwai / Principal Investigator
University of Helsinki, Institute of Biotechnology
Date: 2021-10-01
Language: eng
Number of pages: 13
Belongs to series: Molecules
ISSN: 1420-3049
URI: http://hdl.handle.net/10138/335474
Abstract: An efficient self-cleavable purification tag could be a powerful tool for purifying recom-binant 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 pro-tein-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, Halor-habdus utahensis and applied it to small peptides. Moreover, we described a method for the ami-dation using the same IIST system and demonstrated15N-labeling of the C-terminal amide group of a single domain antibody (VHH).
Subject: 1182 Biochemistry, cell and molecular biology
halophilic intein
NMR
protein splicing
conditional protein splicing
C-terminal amidation
isotopic labeling
peptide purification
purification tag
self-cleaving tag
halophilic intein
NMR
protein splicing
conditional protein splicing
C-terminal amidation
isotopic labeling
peptide purification
purification tag
self-cleaving tag
DNAE INTEIN
STABILIZATION
AMINO
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