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

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

Summary: 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 d...

Full description

Bibliographic Details
Main Authors: Zijun Chen, Jinqing Huang, Jixi Li
Format: Article
Language:English
Published: Elsevier 2022-09-01
Series:STAR Protocols
Subjects:
Molecular Biology
Protein Biochemistry
Protein expression and purification
Structural Biology
X-ray Crystallography
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166722005226
Description
Summary:Summary: 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). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
ISSN:2666-1667

Similar Items