title: Capturing the herpes simplex virus core fusion complex (gB-gH/gL) in an acidic environment.
abstract: Herpes simplex virus (HSV) entry requires the core fusion machinery of gH/gL and gB as well as gD and a gD receptor. When gD binds receptor, it undergoes conformational changes that presumably activate gH/gL, which then activates gB to carry out fusion. gB is a class III viral fusion protein, while gH/gL does not resemble any known viral fusion protein. One hallmark of fusion proteins is their ability to bind lipid membranes. We previously used a liposome coflotation assay to show that truncated soluble gB, but not gH/gL or gD, can associate with liposomes at neutral pH. Here, we show that gH/gL cofloats with liposomes but only when it is incubated with gB at pH 5. When gB mutants with single amino acid changes in the fusion loops (known to inhibit the binding of soluble gB to liposomes) were mixed with gH/gL and liposomes at pH 5, gH/gL failed to cofloat with liposomes. These data suggest that gH/gL does not directly associate with liposomes but instead binds to gB, which then binds to liposomes via its fusion loops. Using monoclonal antibodies, we found that many gH and gL epitopes were altered by low pH, whereas the effect on gB epitopes was more limited. Our liposome data support the concept that low pH triggers conformational changes to both proteins that allow gH/gL to physically interact with gB.
authors.affiliation: Department of Microbiology, University of Pennsylvania, 4010 Locust St., Levy Rm. 233, Philadelphia, PA 19104, USA. tmcairns@biochem.dental.upenn.edu
authors.forename: Tina M
authors.initials: TM
authors.lastname: Cairns
authors.forename: J Charles
authors.initials: JC
authors.lastname: Whitbeck
authors.forename: Huan
authors.initials: H
authors.lastname: Lou
authors.forename: Ekaterina E
authors.initials: EE
authors.lastname: Heldwein
authors.forename: Tirumala K
authors.initials: TK
authors.lastname: Chowdary
authors.forename: Roselyn J
authors.initials: RJ
authors.lastname: Eisenberg
authors.forename: Gary H
authors.initials: GH
authors.lastname: Cohen
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002478:Cells, Cultured; D003460:Crystallization; D005822:Genetic Vectors; D006863:Hydrogen-Ion Concentration; D008081:Liposomes / Q000378:metabolism; D008958:Models, Molecular; D018139:Simplexvirus / Q000378:metabolism*; D018411:Spodoptera; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D008081:Liposomes; D014759:Viral Envelope Proteins; C040495:glycoprotein B, Simplexvirus; C044200:glycoprotein H, herpes simplex virus type 1; C077407:glycoprotein L, Human herpesvirus 1