title: Expression, purification, crystallization and preliminary X-ray analysis of full-length human RIG-I.
abstract: 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 F23, with unit-cell parameters a = b = c = 216.43 Å, α = β = γ = 90°.
authors.affiliation: Department of Physiology, Li Ka Shing Faculty of Medicine, University of Hong Kong, Hong Kong.
authors.forename: Jane
authors.initials: J
authors.lastname: Kwok
authors.affiliation: Department of Microbiology, Li Ka Shing Faculty of Medicine, University of Hong Kong, Hong Kong.
authors.forename: Kenrie P Y
authors.initials: KP
authors.lastname: Hui
authors.affiliation: Division of Structural Biology and Biochemistry, School of Biological Sciences, Nanyang Technological University, Singapore 138673, Singapore.
authors.forename: Julien
authors.initials: J
authors.lastname: Lescar
authors.affiliation: Department of Physiology, Li Ka Shing Faculty of Medicine, University of Hong Kong, Hong Kong.
authors.forename: Masayo
authors.initials: M
authors.lastname: Kotaka
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D000071457:DEAD Box Protein 58; D053487:DEAD-box RNA Helicases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D011487:Protein Conformation; D011971:Receptors, Immunologic
chemical_list: D011971:Receptors, Immunologic; C452820:RIGI protein, human; D000071457:DEAD Box Protein 58; D053487:DEAD-box RNA Helicases
keywords: human RIG-I