Processing of hepatitis B virus surface antigen expressed by recombinant Oka varicella vaccine virus.
We have constructed a recombinant Oka varicella vaccine virus expressing hepatitis B virus (HBV) surface antigen (HBsAg). HBsAg was synthesized as 26K and 30K proteins in infected cells and secreted into the culture supernatant as 30K and 35K proteins. Inhibitors and glycosidase treatments, and pulse-chase labelling experiments, revealed the glycosylation process of HBsAg. The latter was synthesized as a non-glycosylated 26K protein and subjected to N-linked glycosylation to form a 30K protein with high mannose glycans. Three species of dimers composed of 26K and 30K subunits were then formed with disulphide bonds. Both subunits of the dimers were further subjected to O-linked glycosylation and conversion from high mannose glycans to complex glycans followed by sialylation. Three species of dimers composed of 30K and 35K subunits were secreted into the culture supernatant as HBsAg particles. HBsAg was synthesized, glycosylated with both N- and O-linked glycans, sialylated, and then secreted into the culture supernatant within 1 h. These modifications of HBsAg by glycans might stabilize its structure and enhance its immunogenicity as a live HBV vaccine.
Shiraki
K
K
Department of Virology, Toyama Medical and Pharmaceutical University, Japan.
Ochiai
H
H
Matsui
S
S
Aiba
N
N
Yoshida
Y
Y
Okuno
T
T
Yamanishi
K
K
Takahashi
M
M
D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D005822:Genetic Vectors; D006026:Glycoside Hydrolases / Q000378:metabolism; D006031:Glycosylation; D006514:Hepatitis B Surface Antigens / Q000235:genetics / Q000378:metabolism*; D014645:Herpesvirus 3, Human / Q000235:genetics*; D008985:Monensin / Q000494:pharmacology; D011499:Protein Processing, Post-Translational; D014415:Tunicamycin / Q000494:pharmacology
D006514:Hepatitis B Surface Antigens; D014415:Tunicamycin; D008985:Monensin; D006026:Glycoside Hydrolases