title: Potential nectin-1 binding site on herpes simplex virus glycoprotein d.
abstract: Four glycoproteins (gD, gB, gH, and gL) are essential for herpes simplex virus (HSV) entry into cells. An early step of fusion requires gD to bind one of several receptors, such as nectin-1 or herpesvirus entry mediator (HVEM). We hypothesize that a conformational change in gD occurs upon receptor binding that triggers the other glycoproteins to mediate fusion. Comparison of the crystal structures of gD alone and gD bound to HVEM reveals that upon HVEM binding, the gD N terminus transitions from a flexible stretch of residues to a hairpin loop. To address the contribution of this transition to the ability of gD to trigger fusion, we attempted to "lock" the gD N terminus into a looped conformation by engineering a disulfide bond at its N and C termini. The resulting mutant (gD-A3C/Y38C) failed to trigger fusion in the absence of receptor, suggesting that formation of the loop is not the sole fusion trigger. Unexpectedly, although gD-A3C/Y38C bound HVEM, it failed to bind nectin-1. This was due to the key role played by Y38 in interacting with nectin-1. Since tyrosines are often "hot spot" residues at the center of protein-protein interfaces, we mutated residues that surround Y38 on the same face of gD and tested their binding and functional properties. Our results suggest that this region of gD is important for nectin-1 interaction and is distinct from but partially overlaps the site of HVEM binding. Unique gD mutants with altered receptor usage generated in this study may help dissect the roles played by various HSV receptors during infection.
authors.affiliation: Department of Microbiology, School of Dental Medicine, University of Pennsylvania, Philadelphia, USA. connolly@northwestern.edu
authors.forename: Sarah A
authors.initials: SA
authors.lastname: Connolly
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Landsburg
authors.forename: Andrea
authors.initials: A
authors.lastname: Carfi
authors.forename: J Charles
authors.initials: JC
authors.lastname: Whitbeck
authors.forename: Yi
authors.initials: Y
authors.lastname: Zuo
authors.forename: Don C
authors.initials: DC
authors.lastname: Wiley
authors.forename: Gary H
authors.initials: GH
authors.lastname: Cohen
authors.forename: Roselyn J
authors.initials: RJ
authors.lastname: Eisenberg
mesh_terms: D000818:Animals; D001665:Binding Sites; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D018604:Epitope Mapping; D006801:Humans; D008561:Membrane Fusion; D051379:Mice; D000075983:Nectins; D011487:Protein Conformation; D011991:Receptors, Virus / Q000502:physiology; D014759:Viral Envelope Proteins / Q000737:chemistry*
chemical_list: D015815:Cell Adhesion Molecules; C000619010:NECTIN1 protein, human; C000619015:Nectin1 protein, mouse; D000075983:Nectins; D011991:Receptors, Virus; D014759:Viral Envelope Proteins; C039447:glycoprotein D, Human herpesvirus 1