title: Identification of conserved residues in the E2 envelope glycoprotein of the hepatitis C virus that are critical for CD81 binding.
abstract: Hepatitis C virus (HCV) cell entry involves interaction between the viral envelope glycoprotein E2 and the cell surface receptor CD81. Knowledge of conserved E2 determinants important for successful binding will facilitate development of entry inhibitors designed to block this interaction. Previous studies have assigned the CD81 binding function to a number of discontinuous regions of E2. To better define specific residues involved in receptor binding, a panel of mutants of HCV envelope proteins was generated, where conserved residues within putative CD81 binding regions were sequentially mutated to alanine. Mutant proteins were tested for binding to a panel of monoclonal antibodies and CD81 and for their ability to form noncovalent heterodimers and confer infectivity in the retroviral pseudoparticle (HCVpp) assay. Detection by conformation-sensitive monoclonal antibodies indicated that the mutant proteins were correctly folded. Mutant proteins fell into three groups: those that bound CD81 and conferred HCVpp infectivity, those that abrogated both CD81 binding and HCVpp infectivity, and a final group containing mutants that were able to bind CD81 but were noninfectious in the HCVpp assay. Specific amino acids conserved across all genotypes that were critical for CD81 binding were W420, Y527, W529, G530, and D535. These data significantly increase our understanding of the CD81 receptor-E2 binding process.
authors.affiliation: MRC Virology Unit, Institute of Virology, University of Glasgow, UK.
authors.forename: Ania M
authors.initials: AM
authors.lastname: Owsianka
authors.forename: Judith M
authors.initials: JM
authors.lastname: Timms
authors.forename: Alexander W
authors.initials: AW
authors.lastname: Tarr
authors.forename: Richard J P
authors.initials: RJ
authors.lastname: Brown
authors.forename: Timothy P
authors.initials: TP
authors.lastname: Hickling
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Szwejk
authors.forename: Krystyna
authors.initials: K
authors.lastname: Bienkowska-Szewczyk
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Thomson
authors.forename: Arvind H
authors.initials: AH
authors.lastname: Patel
authors.forename: Jonathan K
authors.initials: JK
authors.lastname: Ball
mesh_terms: D000595:Amino Acid Sequence; D015703:Antigens, CD / Q000378:metabolism*; D002460:Cell Line; D017124:Conserved Sequence; D016174:Hepacivirus / Q000235:genetics / Q000472:pathogenicity*; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D011991:Receptors, Virus / Q000378:metabolism*; D060225:Tetraspanin 28; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D015703:Antigens, CD; C554798:CD81 protein, human; D011991:Receptors, Virus; D060225:Tetraspanin 28; D014759:Viral Envelope Proteins; C091015:glycoprotein E2, Hepatitis C virus