title: Role of the propeptide in controlling conformation and assembly state of hepatitis B virus e-antigen.
abstract: Hepatitis B virus "e-antigen" (HBeAg) is thought to be a soluble dimeric protein that is associated with chronic infection. It shares 149 residues with the viral capsid protein "core-antigen" (HBcAg), but has an additional 10-residue, hydrophobic, cysteine-containing amino-terminal propeptide whose presence correlates with physical, serological, and immunological differences between the two proteins. In HBcAg dimers, the subunits pair by forming a four-helix bundle stabilized by an intermolecular disulfide bond. The structure of HBeAg is probably similar but, instead, has two intramolecular disulfide bonds involving the propeptide. To compare the proteins directly and thereby clarify the role of the propeptide, we identified mutations and solution conditions that render both proteins as either soluble dimers or assembled capsids. Thermally induced unfolding monitored by circular dichroism, and electrophoresis of oxidized and reduced dimers, showed that the propeptide has a destabilizing effect and that the intramolecular disulfide bond forms preferentially and blocks the formation of the intermolecular disulfide bond that otherwise stabilizes the dimer. The HBeAg capsids are less regular than the HBcAg capsids; nevertheless, cryo-electron microscopy reconstructions confirm that they are constructed of dimers resembling those of HBcAg capsids. In them, a portion of the propeptide is visible near the dimer interface, suggesting that it intercalates there, consistent with the known formation of a disulfide bond between C(-7) in the propeptide and C61 in the dimer interface. However, this intercalation distorts the dimer into an assembly-reluctant conformation.
authors.affiliation: Protein Expression Laboratory, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Norman R
authors.initials: NR
authors.lastname: Watts
authors.forename: James F
authors.initials: JF
authors.lastname: Conway
authors.forename: Naiqian
authors.initials: N
authors.lastname: Cheng
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Stahl
authors.forename: Alasdair C
authors.initials: AC
authors.lastname: Steven
authors.forename: Paul T
authors.initials: PT
authors.lastname: Wingfield
mesh_terms: D002942:Circular Dichroism; D020285:Cryoelectron Microscopy; D004220:Disulfides / Q000737:chemistry / Q000378:metabolism; D006513:Hepatitis B e Antigens / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D009154:Mutation / Q000235:genetics; D010446:Peptide Fragments / Q000494:pharmacology*; D011487:Protein Conformation
chemical_list: D004220:Disulfides; D006513:Hepatitis B e Antigens; D010446:Peptide Fragments