Facade-Based Bicelles as a New Tool for Production of Active Membrane Proteins in a Cell-Free System.
CFPS
Facade
bicelle
cell-free
detergent
lipid
membrane protein
nanodisc
protein production
synthesis of an active protein
Journal
International journal of molecular sciences
ISSN: 1422-0067
Titre abrégé: Int J Mol Sci
Pays: Switzerland
ID NLM: 101092791
Informations de publication
Date de publication:
03 Oct 2023
03 Oct 2023
Historique:
received:
26
08
2023
revised:
18
09
2023
accepted:
29
09
2023
medline:
2
11
2023
pubmed:
14
10
2023
entrez:
14
10
2023
Statut:
epublish
Résumé
Integral membrane proteins are important components of a cell. Their structural and functional studies require production of milligram amounts of proteins, which nowadays is not a routine process. Cell-free protein synthesis is a prospective approach to resolve this task. However, there are few known membrane mimetics that can be used to synthesize active membrane proteins in high amounts. Here, we present the application of commercially available "Facade" detergents for the production of active rhodopsin. We show that the yield of active protein in lipid bicelles containing Facade-EM, Facade-TEM, and Facade-EPC is several times higher than in the case of conventional bicelles with CHAPS and DHPC and is comparable to the yield in the presence of lipid-protein nanodiscs. Moreover, the effects of the lipid-to-detergent ratio, concentration of detergent in the feeding mixture, and lipid composition of the bicelles on the total, soluble, and active protein yields are discussed. We show that Facade-based bicelles represent a prospective membrane mimetic, available for the production of membrane proteins in a cell-free system.
Identifiants
pubmed: 37834312
pii: ijms241914864
doi: 10.3390/ijms241914864
pmc: PMC10573531
pii:
doi:
Substances chimiques
Membrane Proteins
0
Lipid Bilayers
0
Detergents
0
Micelles
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : Russian Science Foundation
ID : 22-24-00593
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