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Expression, purification, crystallization and preliminary X‐ray analysis of full‐length human RIG‐I
Author(s) -
Kwok Jane,
Hui Kenrie P. Y.,
Lescar Julien,
Kotaka Masayo
Publication year - 2014
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x14000430
Subject(s) - crystallization , x ray , materials science , crystallography , computational biology , biology , chemistry , physics , optics , organic chemistry
The human innate immune system can detect invasion by microbial pathogens through pattern‐recognition receptors that recognize structurally conserved pathogen‐associated molecular patterns. Retinoic acid‐inducible gene I (RIG‐I)‐like helicases (RLHs) are one of the two major families of pattern‐recognition receptors that can detect viral RNA. RIG‐I, belonging to the RLH family, is capable of recognizing intracellular viral RNA from RNA viruses, including influenza virus and Ebola virus. Here, full‐length human RIG‐I (hRIG‐I) was cloned in Escherichia coli and expressed in a recombinant form with a His‐SUMO tag. The protein was purified and crystallized at 291 K using the hanging‐drop vapour‐diffusion method. X‐ray diffraction data were collected to 2.85 Å resolution; the crystal belonged to space group F 23, with unit‐cell parameters a = b = c = 216.43 Å, α = β = γ = 90°.