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Substrate specificities of inteins investigated by QuickDrop‐cassette mutagenesis
Author(s) -
Oeemig Jesper S.,
Beyer Hannes M.,
Aranko A. Sesilja,
Mutanen Justus,
Iwaï Hideo
Publication year - 2020
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1002/1873-3468.13909
Subject(s) - mutagenesis , rna splicing , protein splicing , computational biology , site directed mutagenesis , amino acid , amino acid residue , peptide , biochemistry , biology , chemistry , genetics , peptide sequence , gene , mutation , mutant , rna
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.