Protein purification, crystallization, and structure determination of human DEAD-box RNA helicase DDX21 in different unwinding states

RNA helicase DDX21 plays vital roles in ribosomal RNA processing and the regulation of host innate immunity during virus infection. Here, we describe the optimized protocols for nucleic acid-free protein purification and crystallization of DDX21 in its different unwinding states. Rational design of...

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Autores principales: Chen, Zijun, Huang, Jinqing, Li, Jixi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9420396/
https://www.ncbi.nlm.nih.gov/pubmed/36042885
http://dx.doi.org/10.1016/j.xpro.2022.101642
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author Chen, Zijun
Huang, Jinqing
Li, Jixi
author_facet Chen, Zijun
Huang, Jinqing
Li, Jixi
author_sort Chen, Zijun
collection PubMed
description RNA helicase DDX21 plays vital roles in ribosomal RNA processing and the regulation of host innate immunity during virus infection. Here, we describe the optimized protocols for nucleic acid-free protein purification and crystallization of DDX21 in its different unwinding states. Rational design of the flexible region within the helicase core, and biophysical approach to characterize interactions between DDX21 and RNA, leads to successful crystallization of DDX21. This protocol can be applied to the crystallography of other DExD/H-box RNA helicases. For complete details on the use and execution of this protocol, please refer to Chen et al. (2020).
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spelling pubmed-94203962022-08-29 Protein purification, crystallization, and structure determination of human DEAD-box RNA helicase DDX21 in different unwinding states Chen, Zijun Huang, Jinqing Li, Jixi STAR Protoc Protocol RNA helicase DDX21 plays vital roles in ribosomal RNA processing and the regulation of host innate immunity during virus infection. Here, we describe the optimized protocols for nucleic acid-free protein purification and crystallization of DDX21 in its different unwinding states. Rational design of the flexible region within the helicase core, and biophysical approach to characterize interactions between DDX21 and RNA, leads to successful crystallization of DDX21. This protocol can be applied to the crystallography of other DExD/H-box RNA helicases. For complete details on the use and execution of this protocol, please refer to Chen et al. (2020). Elsevier 2022-08-19 /pmc/articles/PMC9420396/ /pubmed/36042885 http://dx.doi.org/10.1016/j.xpro.2022.101642 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
Chen, Zijun
Huang, Jinqing
Li, Jixi
Protein purification, crystallization, and structure determination of human DEAD-box RNA helicase DDX21 in different unwinding states
title Protein purification, crystallization, and structure determination of human DEAD-box RNA helicase DDX21 in different unwinding states
title_full Protein purification, crystallization, and structure determination of human DEAD-box RNA helicase DDX21 in different unwinding states
title_fullStr Protein purification, crystallization, and structure determination of human DEAD-box RNA helicase DDX21 in different unwinding states
title_full_unstemmed Protein purification, crystallization, and structure determination of human DEAD-box RNA helicase DDX21 in different unwinding states
title_short Protein purification, crystallization, and structure determination of human DEAD-box RNA helicase DDX21 in different unwinding states
title_sort protein purification, crystallization, and structure determination of human dead-box rna helicase ddx21 in different unwinding states
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9420396/
https://www.ncbi.nlm.nih.gov/pubmed/36042885
http://dx.doi.org/10.1016/j.xpro.2022.101642
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