Recombinant expression, purification, and crystallization of the sterile α-motif/histidine-aspartate domain-containing protein from chicken

Protein Expr Purif. 2017 May:133:96-101. doi: 10.1016/j.pep.2016.04.011. Epub 2016 Apr 27.

Abstract

The sterile α-motif and HD domain containing protein 1 (SAMHD1) family is a newly identified protein family, involved in innate immunity restriction. This family possesses a broad-spectrum of antiviral activity. The SAMHD1 family in chicken has not been clearly documented. Here, we expressed chicken SAMHD1 (101-614) fused with a SUMO tag in an Escherichia coli (E. coli) system. For the first time, chicken SAMHD1 (101-614) was found to possess dNTPase cleavage activities in vitro. This suggests that chicken SAMHD1 may be a potential antiviral factor against avian viruses. Through a unique purification method, the purity of the protein as estimated by SDS-PAGE was >95% after a double Ni affinity chromatography and gel filtration purification. Using a sitting-drop vapor-diffusion method, protein crystals were obtained. This study provides some essential method and information for further structure and function determinations of chicken SAMHD1.

Keywords: Enzyme catalysis; Innate immunity; Restriction factor; dNTPase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Avian Proteins* / biosynthesis
  • Avian Proteins* / chemistry
  • Avian Proteins* / genetics
  • Avian Proteins* / isolation & purification
  • Chickens / genetics
  • Chickens / metabolism*
  • Crystallography, X-Ray
  • Monomeric GTP-Binding Proteins* / biosynthesis
  • Monomeric GTP-Binding Proteins* / chemistry
  • Monomeric GTP-Binding Proteins* / genetics
  • Monomeric GTP-Binding Proteins* / isolation & purification
  • Protein Domains
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification

Substances

  • Avian Proteins
  • Recombinant Proteins
  • Monomeric GTP-Binding Proteins