Segmental isotopic labeling of a single-domain globular protein without any refolding step by an asparaginyl endopeptidase.
FEBS Lett
; 591(9): 1285-1294, 2017 05.
Article
de En
| MEDLINE
| ID: mdl-28369872
ABSTRACT
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.
Mots clés
Texte intégral:
1
Collection:
01-internacional
Base de données:
MEDLINE
Sujet principal:
Peptides cycliques
/
Protéines végétales
/
Cysteine endopeptidases
/
Marquage isotopique
Langue:
En
Journal:
FEBS Lett
Année:
2017
Type de document:
Article
Pays d'affiliation:
Finlande