Crystallization and preliminary X-ray analysis of Ebola VP35 interferon inhibitory domain mutant proteins.
VP35 is one of seven structural proteins encoded by the Ebola viral genome and mediates viral replication, nucleocapsid formation and host immune suppression. The C-terminal interferon inhibitory domain (IID) of VP35 is critical for dsRNA binding and interferon inhibition. The wild-type VP35 IID structure revealed several conserved residues that are important for dsRNA binding and interferon antagonism. Here, the expression, purification and crystallization of recombinant Zaire Ebola VP35 IID mutants R312A, K319A/R322A and K339A in space groups P6(1)22, P2(1)2(1)2(1) and P2(1), respectively, are described. Diffraction data were collected using synchrotron sources at the Advanced Light Source and the Advanced Photon Source.
Leung
Daisy W
DW
Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, IA 50011, USA.
Borek
Dominika
D
Farahbakhsh
Mina
M
Ramanan
Parameshwaran
P
Nix
Jay C
JC
Wang
Tianjiao
T
Prins
Kathleen C
KC
Otwinowski
Zbyszek
Z
Honzatko
Richard B
RB
Helgeson
Luke A
LA
Basler
Christopher F
CF
Amarasinghe
Gaya K
GK
D000595:Amino Acid Sequence; D017124:Conserved Sequence; D003460:Crystallization; D018360:Crystallography, X-Ray; D029043:Ebolavirus / Q000737:chemistry* / Q000235:genetics; D007372:Interferons / Q000037:antagonists & inhibitors*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D054334:Viral Regulatory and Accessory Proteins / Q000737:chemistry* / Q000235:genetics
C080980:VP35 protein, filovirus; D054334:Viral Regulatory and Accessory Proteins; D007372:Interferons