[Differences in glycosylation of the E2 protein between virulent Shimen strain and a virulent C-strain of classical swine fever virus].
The E2 envelope glycoprotein of virulent Shimen strain and avirulent C-strain of Classical swine fever virus (CSFV) has 5 and 6 potential glycosylation sites, respectively, and the potential glycosylation site 986N is unique to C-strain. To study the differences in glycosylation between the virus pair, the E2 genes (removing signal sequence and transmembrane anchor regions) of the two strains fused with the melittin signal sequence were expressed in the Sf9 insect cells. The recombinant E2 proteins were secreted into the medium of Sf9 cells in dimer form with different molecular weight (MW). Deglycosylation of the recombinant E2 proteins by endo H and PNGase F showed that the deglycosalated Shimen-E2 and HCLV-E2 have the same MW, indicating that the different MW between Shimen-E2 and HCLV-E2 proteins came from different glycosylation. Site-directed mutagenesis in the potential glycosylation site at 986N demonstrated that the mutated Shimen-E2 protein had the same MW as the wild-type HCLV-E2 protein, while the mutated HCLV-E2 had the same MW as the wild-type Shimen-E2 protein. We suggest that the different MW between Shimen-E2 and HCLV-E2 is resulted from the different glycosylation on 986 N glycosylation site.
Peng
Wu-Ping
WP
Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 150001, China.
Xia
Zhao-He
ZH
Hou
Qiang
Q
Li
Na
N
Sun
Yuan
Y
Tong
Guang-Zhi
GZ
Qiu
Hua-Ji
HJ
D016367:Baculoviridae / Q000235:genetics; D015153:Blotting, Western; D006692:Classical Swine Fever Virus / Q000737:chemistry* / Q000145:classification; D006031:Glycosylation; D008970:Molecular Weight; D009154:Mutation; D011994:Recombinant Proteins / Q000737:chemistry; D014759:Viral Envelope Proteins / Q000737:chemistry*; D014774:Virulence
D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C087086:glycoprotein E2, classical swine fever virus