Phage-displayed peptides mimicking the discontinuous neutralization sites of puumala Hantavirus envelope glycoproteins.
We selected peptide ligands mimicking the surface structure of discontinuous binding sites of Puumala hantavirus-neutralizing monoclonal antibodies from a random 18-amino acid peptide library containing a disulfide bridge in a fixed position and displayed on a filamentous phage. The varying of selection conditions, either by shortening of the association time or by competitive elution with antigen, was crucial for the selection of peptide inserts that could be aligned with the primary sequences of the envelope glycoproteins G1 and G2. Correspondingly, when the envelope glycoprotein sequences were synthesized as overlapping peptides as spots on membrane, the same site in primary structure was found as with phage display, which corroborates the use of the two methods in mapping of conformational epitopes. Also, epitopes reactive with early-phase sera from Puumala virus infection were defined with the pepspot assay in the amino-terminal region of G1. Similarities of the selected phage clones to a monoclonal antibody-escape mutant site and to a linear early-phase epitope were found.
Heiskanen
T
T
Department of Virology, University of Helsinki, Helsinki, FIN-00014, Finland. Tuomas.Heiskanen@helsinki.fi
Lundkvist
A
A
Soliymani
R
R
Koivunen
E
E
Vaheri
A
A
Lankinen
H
H
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000956:Antigens, Viral / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D001667:Binding, Competitive; D003001:Cloning, Molecular; D004220:Disulfides / Q000737:chemistry / Q000378:metabolism; D000939:Epitopes / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D006476:Orthohantavirus / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D018778:Hantavirus Infections / Q000097:blood / Q000276:immunology / Q000821:virology; D006801:Humans; D017912:Inovirus / Q000235:genetics; D018716:Molecular Mimicry; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D019151:Peptide Library; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology; D011487:Protein Conformation; D016415:Sequence Alignment; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D004220:Disulfides; D000939:Epitopes; D019151:Peptide Library; D010455:Peptides; D014759:Viral Envelope Proteins; C104008:glycoprotein G1, Hantavirus; C090197:glycoprotein G2, Hantavirus