title: Synthesis, solution structure analysis and antibody binding of cyclic epitope peptides from glycoprotein D of Herpes simplex virus type I.
abstract: Two cyclic peptides with a thioether bond have been synthesised corresponding to the 9-22 (9LKMADPNRFRGKDL(22)) sequence of glycoprotein D (gD-1) of Herpes simplex virus. The role of the secondary structure in protein-specific monoclonal antibody recognition was investigated. The sequence selected for this study comprises a strongly antigenic site adopting a beta-turn at residues 14Pro-(15)Asn. Thioether bond was formed between the free thiol group of cysteine or homocysteine inserted in position 11 and the chloroacetylated side chain of lysine in position 18. We report here the preparation of cyclic peptides containing Cys or Hcy in position 11, differing only in one methylene group. The linear precursor peptides were synthesised by Boc/Bzl strategy on MBHA resin, and the cyclisation was carried out in alkaline solution. The secondary structure of the peptides was studied by CD, FT-IR and NMR spectroscopy. The CD and FT-IR data have revealed fundamental changes in the solution conformation of the two compounds. The CH(2) group difference significantly resulted in the altered turn structure at the 12Ala and 13Asp as identified by NMR spectroscopy. The antibody binding properties of the cyclopeptides studied by gD-specific monoclonal antibody (A16) in direct and competition enzyme-linked immunosorbent assay (ELISA) were also not the same. We found that peptide LK[HcyADPNRFK]GKDL exhibited higher affinity to Mab A16 than peptide LK[CADPNRFK]GKDL, however, their reactivity was significantly lower compared to the linear ones. Our results clearly show the importance of secondary structure in an antibody binding and demonstrate that even a slight modification of the primary structure dramatically could influence the immune recognition of the synthetic antigens.
authors.affiliation: Research Group of Peptide Chemistry, Eötvös L. University, Hungarian Academy of Sciences, PO Box 32, Budapest 112, H-1518, Hungary.
authors.forename: Gitta
authors.initials: G
authors.lastname: Schlosser
authors.forename: Gábor
authors.initials: G
authors.lastname: Mezo
authors.forename: Róbert
authors.initials: R
authors.lastname: Kiss
authors.forename: Elemér
authors.initials: E
authors.lastname: Vass
authors.forename: Zsuzsa
authors.initials: Z
authors.lastname: Majer
authors.forename: Matty
authors.initials: M
authors.lastname: Feijlbrief
authors.forename: András
authors.initials: A
authors.lastname: Perczel
authors.forename: Szilvia
authors.initials: S
authors.lastname: Bosze
authors.forename: Sytske
authors.initials: S
authors.lastname: Welling-Wester
authors.forename: Ferenc
authors.initials: F
authors.lastname: Hudecz
mesh_terms: D000595:Amino Acid Sequence; D000906:Antibodies / Q000276:immunology*; D002942:Circular Dichroism; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000096:biosynthesis / Q000737:chemistry* / Q000276:immunology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D011487:Protein Conformation; D011522:Protons; D012996:Solutions / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000276:immunology*
chemical_list: D000906:Antibodies; D000939:Epitopes; D011522:Protons; D012996:Solutions; D014759:Viral Envelope Proteins; C039447:glycoprotein D, Human herpesvirus 1