The structure of hepadnaviral core antigens. Identification of free thiols and determination of the disulfide bonding pattern.
A set of wild-type and mutant human, woodchuck, and duck hepatitis viral core proteins have been prepared and used to study the free thiol groups and the disulfide bonding pattern present within the core particle. Human (HBcAg) and woodchuck (WHcAg) core proteins contain 4 cysteine residues, whereas duck (DHcAg) core protein contains a single cysteine residue. Each of the cysteines of HBcAg has been eliminated, either singly or in combinations, by a two-step mutagenesis procedure. All of the proteins were shown to have very similar physical and immunochemical properties. All assemble into essentially identical core particle structures. Therefore disulfide bonds are not essential for core particle formation. No intra-chain disulfide bonds occur. Cys107 is a free thiol buried within the particle structure, whereas Cys48 is present partly as a free sulfhydryl which is exposed at the surface of the particle. Cys61 is always and Cys48 is partly involved in interchain disulfide bonds with the identical residues of another monomer, whereas Cys183 is always involved in a disulfide bond with the Cys183 of a different monomer. WHcAg has the same pattern of bonding, whereas DHcAg lacks any disulfide bonds, and the single free sulfhydryl, Cys153 which is equivalent to Cys107 of HBcAg, is buried.
Zheng
J
J
Department of Biochemistry and Molecular Biophysics, Virginia Commonwealth University, Richmond 23298-0614.
Schödel
F
F
Peterson
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DL
D000595:Amino Acid Sequence; D001483:Base Sequence; D002942:Circular Dichroism; D003545:Cysteine / Q000235:genetics; D004220:Disulfides / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D015613:Hepadnaviridae / Q000276:immunology*; D006512:Hepatitis B Core Antigens / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D007525:Isoelectric Focusing; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010957:Plasmids; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping; D013438:Sulfhydryl Compounds / Q000737:chemistry*
D004220:Disulfides; D006512:Hepatitis B Core Antigens; D013438:Sulfhydryl Compounds; D003545:Cysteine