Simple purification and characterization of soluble and homogenous ABC-F translation factors from Enterococcus faecium.
ABC-F protein
Antibiotic resistance
Heparin chromatography
Ni-NTA chromatography
SUMO
Journal
Protein expression and purification
ISSN: 1096-0279
Titre abrégé: Protein Expr Purif
Pays: United States
ID NLM: 9101496
Informations de publication
Date de publication:
10 2023
10 2023
Historique:
received:
05
12
2022
revised:
09
06
2023
accepted:
16
06
2023
medline:
5
7
2023
pubmed:
25
6
2023
entrez:
24
6
2023
Statut:
ppublish
Résumé
The family of ATP-binding cassette F proteins (ABC-F) is mainly made up of cytosolic proteins involved in regulating protein synthesis, and they are often part of a mechanism that confers resistance to ribosome-targeting antibiotics. The existing literature has emphasized the difficulty of purifying these recombinant proteins because of their very low solubility and stability. Here, we describe a rapid and efficient three-step purification procedure that allows for the production of untagged ABC-F proteins from Enterococcus faecium in the heterologous host Escherichia coli. After four purified ABC-F proteins were produced using this protocol, their biological activities were validated by in vitro experiment. In conclusion, our study provides an invaluable tool for obtaining large amounts of untagged and soluble ABC-F proteins that can then be used for in vitro experiments.
Identifiants
pubmed: 37354924
pii: S1046-5928(23)00096-7
doi: 10.1016/j.pep.2023.106325
pii:
doi:
Substances chimiques
ATP-Binding Cassette Transporters
0
Anti-Bacterial Agents
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
106325Informations de copyright
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