title: Recombinant expression, purification, and crystallization of the sterile α-motif/histidine-aspartate domain-containing protein from chicken.
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.
authors.affiliation: School of Life Sciences, Tianjin University, Tianjin 300072, China.
authors.forename: Yanhong
authors.initials: Y
authors.lastname: Li
authors.affiliation: School of Life Sciences, Tianjin University, Tianjin 300072, China. Electronic address: pengxin_5403@126.com.
authors.forename: Xin
authors.initials: X
authors.lastname: Peng
authors.affiliation: School of Life Sciences, Tianjin University, Tianjin 300072, China. Electronic address: nkqinxiaohong@126.com.
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Qin
mesh_terms: D000818:Animals; D030161:Avian Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D002645:Chickens / Q000235:genetics / Q000378:metabolism*; D018360:Crystallography, X-Ray; D020559:Monomeric GTP-Binding Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D000072417:Protein Domains; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification
chemical_list: D030161:Avian Proteins; D011994:Recombinant Proteins; D020559:Monomeric GTP-Binding Proteins
keywords: Enzyme catalysis; Innate immunity; Restriction factor; dNTPase