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Segmental isotopic labeling of a single‐domain globular protein without any refolding step by an asparaginyl endopeptidase
Author(s) -
Mikula Kornelia M.,
Tascón Igor,
Tommila Jenni J.,
Iwaï Hideo
Publication year - 2017
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 - 10.1002/1873-3468.12640
Subject(s) - chemistry , peptide , endopeptidase , chemical ligation , ligation , recombinant dna , isotopic labeling , protein engineering , native chemical ligation , biochemistry , globular protein , escherichia coli , combinatorial chemistry , microbiology and biotechnology , biology , enzyme , cysteine , organic chemistry , gene
Asparaginyl endopeptidases (AEPs) catalyze head‐to‐tail backbone cyclization of naturally occurring cyclic peptides such as cyclotides, and have become an important peptide‐engineering tool for macrocyclization and peptide ligation. Here, we report efficient protein ligation in trans by mimicking efficient backbone cyclization by an AEP without any excess of reactants. We demonstrate a practical application of segmental isotopic labeling for NMR studies of a single‐domain globular protein without any refolding step using the recombinant AEP prepared from Escherichia coli . This simple protein ligation approach using an AEP could be applied for incorporation of various biophysical probes into proteins as well as post‐translational production of full‐length proteins.