Crystallization and preliminary X-ray diffraction analysis of the carbohydrate-recognizing domain (VP8*) of bovine rotavirus strain NCDV.
The infectivity of rotavirus is dramatically enhanced by proteolytic cleavage of its outer layer VP4 spike protein into two functional domains, VP8* and VP5*. The carbohydrate-recognizing domain VP8* is proposed to bind sialic acid-containing host cell-surface glycans and this is followed by a series of subsequent virus-cell interactions. Live attenuated human and bovine rotavirus vaccine candidates for the prevention of gastroenteritis have been derived from bovine rotavirus strain NCDV. The NCDV VP8*(64-224) was overexpressed, purified to homogeneity and crystallized in the presence of an N-acetylneuraminic acid derivative. X-ray diffraction data were collected to a resolution of 2.0 A and the crystallographic structure of NCDV VP8*(64-224) was determined by molecular replacement.
Yu
Xing
X
Institute for Glycomics, Gold Coast Campus, Griffith University, Queensland 4222, Australia.
Guillon
Annabel
A
Szyczew
Alex J
AJ
Kiefel
Milton J
MJ
Coulson
Barbara S
BS
von Itzstein
Mark
M
Blanchard
Helen
H
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D002241:Carbohydrates / Q000737:chemistry*; D002417:Cattle; D003460:Crystallization; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D012401:Rotavirus / Q000737:chemistry* / Q000235:genetics; D013912:Threonine / Q000378:metabolism; D014170:Transformation, Genetic; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D014961:X-Ray Diffraction / Q000379:methods*
D002241:Carbohydrates; D016601:RNA-Binding Proteins; D017361:Viral Nonstructural Proteins; C079234:NS35 protein, rotavirus; D013912:Threonine