Methods for Heterologous Overproduction of Fe-S Proteins.
Anaerobic
Cluster
DPOR
Electron transfer
Fe-S protein expression
[4Fe-4S]
[Fe-S]
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2021
2021
Historique:
entrez:
22
7
2021
pubmed:
23
7
2021
medline:
7
1
2022
Statut:
ppublish
Résumé
Proteins carrying iron-sulfur ([Fe-S]) clusters are critical to the basic metabolism of all organisms. Structural and biochemical investigations of many such [Fe-S] cluster proteins depend on recombinant overproduction using heterologous bacterial hosts such as Escherichia coli . Here, we describe a detailed procedure for the overproduction and purification of two oxygen-sensitive component proteins of the dark-operative protochlorophyllide oxidoreductase (DPOR) complex. The method relies on an engineered Escherichia coli cell line carrying a correction in its genome to restore the loss of a key [Fe-S] cluster biogenesis pathway. The method can also be potentially adapted for the overproduction of other Fe-S proteins.
Identifiants
pubmed: 34292544
doi: 10.1007/978-1-0716-1605-5_4
doi:
Substances chimiques
Iron-Sulfur Proteins
0
Sulfur
70FD1KFU70
Iron
E1UOL152H7
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
69-78Informations de copyright
© 2021. Springer Science+Business Media, LLC, part of Springer Nature.
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