Expression, purification, crystallization and preliminary X-ray analysis of full-length human RIG-I

Acta Crystallogr F Struct Biol Commun. 2014 Feb;70(Pt 2):248-51. doi: 10.1107/S2053230X14000430. Epub 2014 Jan 22.

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°.

Keywords: human RIG-I.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Crystallization
  • Crystallography, X-Ray / methods*
  • DEAD Box Protein 58
  • DEAD-box RNA Helicases / chemistry
  • DEAD-box RNA Helicases / genetics*
  • DEAD-box RNA Helicases / isolation & purification
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Protein Conformation
  • Receptors, Immunologic

Substances

  • Receptors, Immunologic
  • RIGI protein, human
  • DEAD Box Protein 58
  • DEAD-box RNA Helicases