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
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author Thakur, Naveen
Wei, Shushu
Ray, Arka Prabha
Lamichhane, Rajan
Eddy, Matthew T.
author_facet Thakur, Naveen
Wei, Shushu
Ray, Arka Prabha
Lamichhane, Rajan
Eddy, Matthew T.
author_sort Thakur, Naveen
collection PubMed
description 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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spelling pubmed-92936692022-07-20 Production of human A(2A)AR in lipid nanodiscs for (19)F-NMR and single-molecule fluorescence spectroscopy Thakur, Naveen Wei, Shushu Ray, Arka Prabha Lamichhane, Rajan Eddy, Matthew T. STAR Protoc Protocol 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). Elsevier 2022-07-13 /pmc/articles/PMC9293669/ /pubmed/35839771 http://dx.doi.org/10.1016/j.xpro.2022.101535 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Thakur, Naveen
Wei, Shushu
Ray, Arka Prabha
Lamichhane, Rajan
Eddy, Matthew T.
Production of human A(2A)AR in lipid nanodiscs for (19)F-NMR and single-molecule fluorescence spectroscopy
title Production of human A(2A)AR in lipid nanodiscs for (19)F-NMR and single-molecule fluorescence spectroscopy
title_full Production of human A(2A)AR in lipid nanodiscs for (19)F-NMR and single-molecule fluorescence spectroscopy
title_fullStr Production of human A(2A)AR in lipid nanodiscs for (19)F-NMR and single-molecule fluorescence spectroscopy
title_full_unstemmed Production of human A(2A)AR in lipid nanodiscs for (19)F-NMR and single-molecule fluorescence spectroscopy
title_short Production of human A(2A)AR in lipid nanodiscs for (19)F-NMR and single-molecule fluorescence spectroscopy
title_sort production of human a(2a)ar in lipid nanodiscs for (19)f-nmr and single-molecule fluorescence spectroscopy
topic Protocol
url 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
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