A protease-sensitive hinge linking the two domains of the hepatitis B virus core protein is exposed on the viral capsid surface.
Core particles of hepatitis B virus are assembled from dimers of a single 185-residue (subtype adw) viral capsid or core protein (p21.5) which possesses two distinct domains: residues 1 to 144 form a minimal capsid assembly domain, and the arginine-rich, carboxyl-terminal residues 150 to 185 form a protamine-like domain that mediates nucleic acid binding. Little is known about the topography of the p21.5 polypeptide within either the p21.5 capsids or dimers. Here, using site-specific proteases and monoclonal antibodies, we have defined the accessibility of p21.5 residues in dimers and capsids assembled from wild-type and mutant hepatitis B virus core proteins in Xenopus oocytes and in vitro. The data reveal the protamine region to be accessible to external reagents in p21.5 dimers but largely cryptic in wild-type capsids. Strikingly, in capsids the only protease target region was a 9-residue peptide covering p21.5 residues Glu-145 to Asp-153, which falls largely between the two core protein domains. By analogy with protease-sensitive interdomain regions in other proteins, we propose that this peptide constitutes a hinge between the assembly and nucleic acid binding domains of p21.5. We further found that deletion or replacement of the terminal Cys-185 residue greatly increased surface exposure of the protamine tails in capsids, suggesting that a known disulfide linkage involving this residue tethers the protamine region inside the core particles. We propose that disruption of this disulfide linkage allows the protamine region to appear transiently on the surface of the core particle.
Seifer
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Hormone Research Institute, University of California at San Francisco 94143-0534.
Standring
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D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D002213:Capsid / Q000648:ultrastructure*; D010450:Endopeptidases / Q000494:pharmacology; D000939:Epitopes; D006512:Hepatitis B Core Antigens / Q000648:ultrastructure*; D006515:Hepatitis B virus / Q000648:ultrastructure*; D008969:Molecular Sequence Data; D011479:Protamines / Q000737:chemistry
D000911:Antibodies, Monoclonal; D000939:Epitopes; D006512:Hepatitis B Core Antigens; D011479:Protamines; D010450:Endopeptidases