Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of the VP8* carbohydrate-binding protein of the human rotavirus strain Wa.
Rotaviruses exhibit host-specificity and the first crystallographic information on a rotavirus strain that infects humans is reported here. Recognition and attachment to host cells, leading to invasion and infection, is critically linked to the function of the outer capsid spike protein of the rotavirus particle. In some strains the VP8* component of the spike protein is implicated in recognition and binding of sialic-acid-containing cell-surface carbohydrates, thereby enabling infection by the virus. The cloning, expression, purification, crystallization and initial X-ray diffraction analysis of the VP8* core from human Wa rotavirus is reported. Two crystal forms (trigonal P3(2)21 and monoclinic P2(1)) have been obtained and X-ray diffraction data have been collected, enabling determination of the VP8*(64-223) structure by molecular replacement.
Kraschnefski
Mark J
MJ
Institute for Glycomics, Griffith University, PMB 50 Gold Coast Mail Centre, Queensland 9726, Australia.
Scott
Stacy A
SA
Holloway
Gavan
G
Coulson
Barbara S
BS
von Itzstein
Mark
M
Blanchard
Helen
H
D002241:Carbohydrates / Q000737:chemistry*; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D003460:Crystallization; D017931:DNA Primers / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D019158:N-Acetylneuraminic Acid / Q000737:chemistry; D011485:Protein Binding; D012367:RNA, Viral / Q000737:chemistry; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics*; D012401:Rotavirus / Q000235:genetics*; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000235:genetics*; D014961:X-Ray Diffraction
D002241:Carbohydrates; D017931:DNA Primers; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D017361:Viral Nonstructural Proteins; C079234:NS35 protein, rotavirus; D019158:N-Acetylneuraminic Acid