Multiple peptides homologous to herpes simplex virus type 1 glycoprotein B inhibit viral infection.
The 773-residue ectodomain of the herpes simplex virus type 1 (HSV-1) glycoprotein B (gB) has been resistant to the use of mutagenic strategies because the majority of the induced mutations result in defective proteins. As an alternative strategy for the identification of functionally important regions and novel inhibitors of infection, we prepared a library of overlapping peptides homologous to the ectodomain of gB and screened for the ability of the peptides to block infection. Seven of 138 15-mer peptides inhibited infection by more than 50% at a concentration of 100 microM. Three peptides (gB94, gB122, and gB131) with 50% effective concentrations (EC(50)s) below 20 microM were selected for further studies. The gB131 peptide (residues 681 to 695 in HSV-1 gB [gB-1]) was a specific entry inhibitor (EC(50), approximately 12 microM). The gB122 peptide (residues 636 to 650 in gB-1) blocked viral entry (EC(50), approximately 18 microM), protected cells from infection (EC(50), approximately 72 microM), and inactivated virions in solution (EC(50), approximately 138 microM). We were unable to discern the step or steps inhibited by the gB94 peptide, which is homologous to residues 496 to 510 in gB-1. Substitution of a tyrosine in the gB122 peptide (Y640 in full-length gB-1) reduced the antiviral activity eightfold, suggesting that this residue is critical for inhibition. This peptide-based strategy could lead to the identification of functionally important regions of gB or other membrane proteins and identify novel inhibitors of HSV-1 entry.
Akkarawongsa
Radeekorn
R
Department of Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
Pocaro
Nina E
NE
Case
Gary
G
Kolb
Aaron W
AW
Brandt
Curtis R
CR
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D001665:Binding Sites; D002522:Chlorocebus aethiops; D004220:Disulfides / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D006561:Herpes Simplex / Q000517:prevention & control*; D018259:Herpesvirus 1, Human / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D057927:Hydrophobic and Hydrophilic Interactions; D020128:Inhibitory Concentration 50; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000276:immunology / Q000494:pharmacology; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D014443:Tyrosine / Q000378:metabolism; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D014771:Virion / Q000502:physiology
D004220:Disulfides; D010455:Peptides; D014759:Viral Envelope Proteins; C040495:glycoprotein B, Simplexvirus; D014443:Tyrosine