Identification of linear heparin-binding peptides derived from human respiratory syncytial virus fusion glycoprotein that inhibit infectivity.
It has been shown previously that the fusion glycoprotein of human respiratory syncytial virus (RSV-F) interacts with cellular heparan sulfate. Synthetic overlapping peptides derived from the F-protein sequence of RSV subtype A (strain A2) were tested for their ability to bind heparin using heparin-agarose affinity chromatography (HAAC). This evaluation identified 15 peptides representing eight linear heparin-binding domains (HBDs) located within F1 and F2 and spanning the protease cleavage activation site. All peptides bound to Vero and A549 cells, and binding was inhibited by soluble heparins and diminished by either enzymatic treatment to remove cell surface glycosaminoglycans or by treatment with sodium chlorate to decrease cellular sulfation. RSV-F HBD peptides were less likely to bind to glycosaminoglycan-deficient CHO-745 cells than parental CHO-K1 cells that express these molecules. Three RSV-F HBD peptides (F16, F26, and F55) inhibited virus infectivity; two of these peptides (F16 and F55) inhibited binding of virus to Vero cells, while the third (F26) did not. These studies provided evidence that two of the linear HBDs mapped by peptides F16 and F55 may mediate one of the first steps in the attachment of virus to cells while the third, F26, inhibited infectivity at a postattachment step, suggesting that interactions with cell surface glycosaminoglycans may play a role in infectivity of some RSV strains.
Crim
Roberta L
RL
Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892, USA.
Audet
Susette A
SA
Feldman
Steven A
SA
Mostowski
Howard S
HS
Beeler
Judy A
JA
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D016466:CHO Cells; D002704:Chlorates / Q000494:pharmacology; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003412:Cricetulus; D015683:Gene Products, gag / Q000235:genetics / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism; D006493:Heparin / Q000378:metabolism*; D006801:Humans; D008190:Lyases / Q000494:pharmacology; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D010455:Peptides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D018113:Respiratory Syncytial Virus, Human / Q000187:drug effects / Q000378:metabolism / Q000472:pathogenicity*; D014709:Vero Cells; D014760:Viral Fusion Proteins / Q000737:chemistry* / Q000378:metabolism
D002704:Chlorates; D015683:Gene Products, gag; D006023:Glycoproteins; D010455:Peptides; D014760:Viral Fusion Proteins; D006493:Heparin; D008190:Lyases; C032706:sodium chlorate