Incorporating thioamides into proteins by native chemical ligation.
Calmodulin
GB1
Native chemical ligation
Semi-synthesis
Thioamide
α-Synuclein
Journal
Methods in enzymology
ISSN: 1557-7988
Titre abrégé: Methods Enzymol
Pays: United States
ID NLM: 0212271
Informations de publication
Date de publication:
2021
2021
Historique:
entrez:
30
7
2021
pubmed:
31
7
2021
medline:
14
8
2021
Statut:
ppublish
Résumé
The thioamide is a versatile replacement of the peptide backbone with altered hydrogen bonding and conformational preferences, as well the ability participate in energy and electron transfer processes. Semi-synthetic incorporation of a thioamide into a protein can be used to study protein folding or protein/protein interactions using these properties. Semi-synthesis also provides the opportunity to study the role of thioamides in natural proteins. Here we outline the semi-synthesis of a model protein, the B1 domain of protein G (GB1) with a thioamide at the N-terminus or the C-terminus. The thioamide is synthetically incorporated into a fragment by solid-phase peptide synthesis, whereas the remainder of the protein is recombinantly expressed. Then, the two fragments are joined by native chemical ligation. The explicit protocol for GB1 synthesis is accompanied by examples of applications with GB1 and other proteins in structural biology and protein misfolding studies.
Identifiants
pubmed: 34325791
pii: S0076-6879(21)00146-4
doi: 10.1016/bs.mie.2021.04.011
pmc: PMC8617429
mid: NIHMS1755857
pii:
doi:
Substances chimiques
Peptides
0
Proteins
0
Thioamides
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
295-339Subventions
Organisme : NIGMS NIH HHS
ID : T32 GM008275
Pays : United States
Organisme : NIGMS NIH HHS
ID : T32 GM132039
Pays : United States
Informations de copyright
Copyright © 2021 Elsevier Inc. All rights reserved.
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