Relevance of cysteine residues for biosynthesis and antigenicity of human hepatitis B virus e protein.
The mature form of the secretory core protein (HBe protein) of human hepatitis B virus contains four cysteines which are located at amino acid positions -7, 48, 61, and 107 relative to the HBc start methionine. In addition, there is a cysteine, Cys-183, located in the C-terminal domain of the HBe precursor, which is cleaved during HBe maturation. Here, the significance of these cysteines for biosynthesis and antigenicity of the HBe protein was examined. The cysteines at positions -7 and 61 were found to be crucial for HBe biosynthesis. As has already been described, if the Cys at position -7 is mutated, disulfide-linked HBe homodimers which have both HBe antigenicity and HBc antigenicity are expressed. Here we show that these dimers are due to Cys-61-Cys-61 disulfide bridges which are formed only if the Cys at position -7 is not present. In the wild-type protein, this dimerization appears to be inhibited by formation of intramolecular disulfide bridges between the Cys at -7 and one of the internal cysteines. Moreover, Cys-61 is important for HBe biosynthesis in general since mutation of this amino acid results in production of HBe proteins which are either only poorly secreted or possess a different antigenicity.
Wasenauer
G
G
Department of Virology, University of Ulm, Germany.
Köck
J
J
Schlicht
H J
HJ
D000911:Antibodies, Monoclonal; D000917:Antibody Formation; D002478:Cells, Cultured; D003545:Cysteine; D004220:Disulfides / Q000378:metabolism; D005814:Genes, Viral / Q000235:genetics*; D006510:Hepatitis B Antibodies / Q000276:immunology*; D006513:Hepatitis B e Antigens / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D006515:Hepatitis B virus / Q000235:genetics / Q000276:immunology*; D006801:Humans; D008099:Liver / Q000166:cytology; D016297:Mutagenesis, Site-Directed; D016133:Polymerase Chain Reaction; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D011933:Reagent Kits, Diagnostic; D013329:Structure-Activity Relationship; D014616:Vaccinia virus / Q000235:genetics
D000911:Antibodies, Monoclonal; D004220:Disulfides; D006510:Hepatitis B Antibodies; D006513:Hepatitis B e Antigens; D011933:Reagent Kits, Diagnostic; D003545:Cysteine