Functional fine-mapping and molecular modeling of a conserved loop epitope of the measles virus hemagglutinin protein.
Neutralizing and protective monoclonal antibodies (mAbs) were used to fine-map the highly conserved hemagglutinin noose epitope (H379-410, HNE) of the measles virus. Short peptides mimicking this epitope were previously shown to induce virus-neutralizing antibodies [El Kasmi et al. (2000) J. Gen. Virol.81, 729-735]. The epitope contains three cysteine residues, two of which (Cys386 and Cys394) form a disulfide bridge critical for antibody binding. Substitution and truncation analogues revealed four residues critical for binding (Lys387, Gly388, Gln391 and Glu395) and suggested the binding motif X7C[KR]GX[AINQ]QX2CEX5 for three distinct protective mAbs. This motif was found in more than 90% of the wild-type viruses. An independent molecular model of the core epitope predicted an amphiphilic loop displaying a remarkably stable and rigid loop conformation. The three hydrophilic contact residues Lys387, Gln391 and Glu395 pointed on the virus towards the solvent-exposed side of the planar loop and the permissive hydrophobic residues Ile390, Ala392 and Leu393 towards the solvent-hidden side of the loop, precluding antibody binding. The high affinity (Kd = 7.60 nm) of the mAb BH216 for the peptide suggests a high structural resemblance of the peptide with the natural epitope and indicates that most interactions with the protein are also contributed by the peptide. Improved peptides designed on the basis of these findings induced sera that crossreacted with the native measles virus hemagglutinin protein, providing important information about a lead structure for the design of more stable antigens of a synthetic or recombinant subunit vaccine.
Pütz
Mike M
MM
Department of Immunology, Laboratoire National de Santé, Luxembourg; Fakultät für Chemie und Pharmazie, Universität Tübingen, Germany.
Hoebeke
Johan
J
Ammerlaan
Wim
W
Schneider
Serge
S
Muller
Claude P
CP
D020816:Amino Acid Motifs / Q000276:immunology / Q000502:physiology; D000595:Amino Acid Sequence; D000918:Antibody Specificity; D015374:Biosensing Techniques; D017124:Conserved Sequence; D004220:Disulfides; D000939:Epitopes / Q000737:chemistry*; D005434:Flow Cytometry; D006389:Hemagglutinins, Viral / Q000737:chemistry* / Q000276:immunology; D008459:Measles virus / Q000737:chemistry* / Q000276:immunology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D011485:Protein Binding / Q000502:physiology; D020033:Protein Isoforms / Q000737:chemistry / Q000276:immunology; D017386:Sequence Homology, Amino Acid; D020349:Surface Plasmon Resonance; D014407:Tumor Cells, Cultured
D004220:Disulfides; D000939:Epitopes; D006389:Hemagglutinins, Viral; D020033:Protein Isoforms