In Vitro Reconstitution Platforms of Mammalian Cell-Free Expressed Membrane Proteins

Autor:	Hossein Moghimianavval, Yen-Yu Hsu, Alessandro Groaz, Allen P. Liu
Rok vydání:	2022
Předmět:	Cell Membrane Lipid Bilayers Animals Membrane Proteins Unilamellar Liposomes Article
Zdroj:	Methods Mol Biol Cell-Free Gene Expression ISBN: 9781071619971
ISSN:	1940-6029
Popis:	Membrane proteins are essential components in cell membranes and enable cells to communicate with their outside environment and to carry out intracellular signaling. Functional reconstitution of complex membrane proteins using cell-free expression (CFE) systems has been proved to be challenging mainly due to the lack of necessary machinery for proper folding and translocation of nascent membrane proteins and their delivery to the supplied synthetic bilayers. Here, we provide protocols for detergent-free, cell-free reconstitution of functional membrane proteins using HeLa-based CFE system and outline assays for studying their membrane insertion, topology, and their orientation upon incorporation into the supported lipid bilayers or bilayers of giant unilamellar vesicles as well as methods to isolate functional translocated cell-free produced membrane proteins.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::41a15174889788ef47f75c214d28e016 https://pubmed.ncbi.nlm.nih.gov/34985740 Zobrazit plný text záznamu