title: Mutational analysis of the major heparan sulfate-binding domain of herpes simplex virus type 1 glycoprotein C.
abstract: Heparan sulfate (HS) has been identified as a receptor molecule for numerous microbial pathogens, including herpes simplex virus type 1 (HSV-1). To further define the major HS-binding domain of the HSV-1 attachment protein, i.e. glycoprotein C (gC), virus mutants carrying alterations of either two neighbouring basic amino acid residues or a single hydrophobic amino acid residue within the N-terminal domain of the protein (residues 26-227) were constructed. In addition, a mutant lacking the Asn148 glycosylation site was included in the study. Binding of purified mutated gC proteins to isolated HS chains showed that viruses with mutations at residues Arg(129,130), Ile142, Arg(143,145), Arg(145,147), Arg(151,155) and Arg(155,160) had significantly impaired HS binding, in contrast to the other mutations, including Asn148. Impairment of the HS-binding activity of gC by these mutations had profound consequences for virus attachment and infection of cells in which amounts of HS exposed on the cell surface had been reduced. It is suggested that basic and hydrophobic residues localized at the Cys127-Cys144 loop of HSV-1 gC constitute a major HS-binding domain, with the most active amino acids situated near the C-terminal region of the two cysteines.
authors.affiliation: Department of Virology, Göteborg University, Guldhedsgatan 10b, S-413 46 Göteborg, Sweden1.
authors.forename: Kristina
authors.initials: K
authors.lastname: Mårdberg
authors.affiliation: Department of Virology, Göteborg University, Guldhedsgatan 10b, S-413 46 Göteborg, Sweden1.
authors.forename: Edward
authors.initials: E
authors.lastname: Trybala
authors.affiliation: Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA2.
authors.forename: Joseph C
authors.initials: JC
authors.lastname: Glorioso
authors.affiliation: Department of Virology, Göteborg University, Guldhedsgatan 10b, S-413 46 Göteborg, Sweden1.
authors.forename: Tomas
authors.initials: T
authors.lastname: Bergström
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine / Q000235:genetics; D001216:Asparagine / Q000235:genetics; D002460:Cell Line; D005057:Eukaryotic Cells / Q000187:drug effects / Q000378:metabolism / Q000821:virology; D019763:Heparin Lyase / Q000494:pharmacology; D006497:Heparan Sulfate / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011991:Receptors, Virus / Q000378:metabolism*; D018139:Simplexvirus / Q000235:genetics* / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000378:metabolism; D014779:Virus Replication
chemical_list: D001120:Arginine; D001216:Asparagine; D019763:Heparin Lyase; D006497:Heparan Sulfate; D011991:Receptors, Virus; D014759:Viral Envelope Proteins; C038535:glycoprotein gC, herpes simplex virus type 1