Expression, purification, crystallization and preliminary X-ray diffraction analysis of the VP8* sialic acid-binding domain of porcine rotavirus strain OSU.
The rotavirus outer capsid spike protein VP4 is utilized in the process of rotavirus attachment to and membrane penetration of host cells. VP4 is cleaved by trypsin into two domains: VP8* and VP5*. The VP8* domain is implicated in initial interaction with sialic acid-containing cell-surface carbohydrates and triggers subsequent virus invasion. The VP8* domain from porcine OSU rotavirus was cloned and expressed in Escherichia coli. Different crystal forms (orthorhombic P2(1)2(1)2(1) and tetragonal P4(1)2(1)2) were harvested from two distinct crystallization conditions. Diffraction data have been collected to 2.65 and 2.2 A resolution and the VP8*(65-224) structure was determined by molecular replacement.
Zhang
Yang-De
YD
National Hepatobiliary and Enteric Surgery Research Center of The Ministry of Health, Xiangya Hospital, Central South University, Hunan Province, People's Republic of China. zhangyd1960@yahoo.com.cn
Li
Hao
H
Liu
Hui
H
Pan
Yi-Feng
YF
D000818:Animals; D003001:Cloning, Molecular; D003460:Crystallization; D017434:Protein Structure, Tertiary; D016601:RNA-Binding Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D012401:Rotavirus / Q000737:chemistry* / Q000235:genetics; D013552:Swine; D017361:Viral Nonstructural Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D014961:X-Ray Diffraction
D016601:RNA-Binding Proteins; D017361:Viral Nonstructural Proteins; C079234:NS35 protein, rotavirus