Expression, purification, crystallization and preliminary X-ray studies of the Ebola VP35 interferon inhibitory domain.
Ebola VP35 is a multifunctional protein that is important for host immune suppression and pathogenesis. VP35 contains an N-terminal oligomerization domain and a C-terminal interferon inhibitory domain (IID). Mutations within the VP35 IID result in loss of host immune suppression. Here, efforts to crystallize recombinantly overexpressed VP35 IID that was purified from Escherichia coli are described. Native and selenomethionine-labeled crystals belonging to the orthorhombic space group P2(1)2(1)2(1) were obtained by the hanging-drop vapor-diffusion method and diffraction data were collected at the ALS synchrotron.
Leung
Daisy W
DW
Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, IA 50011, USA.
Ginder
Nathaniel D
ND
Nix
Jay C
JC
Basler
Christopher F
CF
Honzatko
Richard B
RB
Amarasinghe
Gaya K
GK
D003460:Crystallization; D029043:Ebolavirus / Q000737:chemistry* / Q000472:pathogenicity*; D015967:Gene Expression Regulation, Viral / Q000502:physiology*; D007372:Interferons / Q000037:antagonists & inhibitors* / Q000737:chemistry; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins / Q000096:biosynthesis / Q000737:chemistry* / Q000502:physiology*; D017434:Protein Structure, Tertiary / Q000502:physiology; D014758:Viral Core Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000502:physiology*; D054334:Viral Regulatory and Accessory Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D014774:Virulence; D014961:X-Ray Diffraction
D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D014758:Viral Core Proteins; D054334:Viral Regulatory and Accessory Proteins; C084818:nucleoprotein VP35, Ebola virus; D007372:Interferons