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Preparation of co-crystals of human PPARα-LBD and ligand for high-resolution X-ray crystallography

Peroxisome proliferator-activated receptors (PPARs) are nuclear receptor-type transcription factors with three subtypes (α, δ, and γ) that regulate cell differentiation and metabolism. Co-crystals of human PPARα-ligand-binding domain (LBD)-PPARα ligand for X-ray crystallography have been difficult t...

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Detalles Bibliográficos
Autores principales: Kamata, Shotaro, Oyama, Takuji, Ishii, Isao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7933539/
https://www.ncbi.nlm.nih.gov/pubmed/33718889
http://dx.doi.org/10.1016/j.xpro.2021.100364
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author Kamata, Shotaro
Oyama, Takuji
Ishii, Isao
author_facet Kamata, Shotaro
Oyama, Takuji
Ishii, Isao
author_sort Kamata, Shotaro
collection PubMed
description Peroxisome proliferator-activated receptors (PPARs) are nuclear receptor-type transcription factors with three subtypes (α, δ, and γ) that regulate cell differentiation and metabolism. Co-crystals of human PPARα-ligand-binding domain (LBD)-PPARα ligand for X-ray crystallography have been difficult to obtain. Recombinant human PPARα-LBD proteins contain intrinsic fatty acids (iFAs of Escherichia coli origin) and may be unstable without ligands during crystallization. To circumvent these limitations, we have successfully applied various crystallization techniques, including co-crystallization, cross-seeding, soaking, delipidation, and coactivator peptide supplementation. For complete details on the use and execution of this protocol, please refer to Kamata et al. (2020).
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spelling pubmed-79335392021-03-12 Preparation of co-crystals of human PPARα-LBD and ligand for high-resolution X-ray crystallography Kamata, Shotaro Oyama, Takuji Ishii, Isao STAR Protoc Protocol Peroxisome proliferator-activated receptors (PPARs) are nuclear receptor-type transcription factors with three subtypes (α, δ, and γ) that regulate cell differentiation and metabolism. Co-crystals of human PPARα-ligand-binding domain (LBD)-PPARα ligand for X-ray crystallography have been difficult to obtain. Recombinant human PPARα-LBD proteins contain intrinsic fatty acids (iFAs of Escherichia coli origin) and may be unstable without ligands during crystallization. To circumvent these limitations, we have successfully applied various crystallization techniques, including co-crystallization, cross-seeding, soaking, delipidation, and coactivator peptide supplementation. For complete details on the use and execution of this protocol, please refer to Kamata et al. (2020). Elsevier 2021-02-26 /pmc/articles/PMC7933539/ /pubmed/33718889 http://dx.doi.org/10.1016/j.xpro.2021.100364 Text en © 2021 The Author(s) http://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
Kamata, Shotaro
Oyama, Takuji
Ishii, Isao
Preparation of co-crystals of human PPARα-LBD and ligand for high-resolution X-ray crystallography
title Preparation of co-crystals of human PPARα-LBD and ligand for high-resolution X-ray crystallography
title_full Preparation of co-crystals of human PPARα-LBD and ligand for high-resolution X-ray crystallography
title_fullStr Preparation of co-crystals of human PPARα-LBD and ligand for high-resolution X-ray crystallography
title_full_unstemmed Preparation of co-crystals of human PPARα-LBD and ligand for high-resolution X-ray crystallography
title_short Preparation of co-crystals of human PPARα-LBD and ligand for high-resolution X-ray crystallography
title_sort preparation of co-crystals of human pparα-lbd and ligand for high-resolution x-ray crystallography
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7933539/
https://www.ncbi.nlm.nih.gov/pubmed/33718889
http://dx.doi.org/10.1016/j.xpro.2021.100364
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