Production of human A(2A)AR in lipid nanodiscs for (19)F-NMR and single-molecule fluorescence spectroscopy

We describe production of the human A(2A) adenosine receptor (A(2A)AR), a class A G protein-coupled receptor (GPCR) for (19)F-NMR and single-molecule fluorescence (SMF) spectroscopy. We explain in detail steps shared between the two sample preparation strategies, including expression and isolation o...

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Detalles Bibliográficos
Autores principales: Thakur, Naveen, Wei, Shushu, Ray, Arka Prabha, Lamichhane, Rajan, Eddy, Matthew T.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9293669/
https://www.ncbi.nlm.nih.gov/pubmed/35839771
http://dx.doi.org/10.1016/j.xpro.2022.101535
Descripción
Sumario:We describe production of the human A(2A) adenosine receptor (A(2A)AR), a class A G protein-coupled receptor (GPCR) for (19)F-NMR and single-molecule fluorescence (SMF) spectroscopy. We explain in detail steps shared between the two sample preparation strategies, including expression and isolation of A(2A)AR and assembly of A(2A)AR in lipid nanodiscs and procedures for incorporation of either (19)F-NMR or fluorescence probes. Protocols for SMF experiments include sample setup, data acquisition, data processing, and error analysis. For complete details on the use and execution of this protocol, please refer to Wei et al. (2022) and Sušac et al. (2018).

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