Monoclonal antibody screening of a phage-displayed random peptide library reveals mimotopes of chemokine receptor CCR5: Implications for the tertiary structure of the receptor and for an n-terminal binding site for HIV-1 gp120
Königs, C., Rowley, M. J., Thompson, P., Myers, M. A., Scealy, M., Davies, Janet M., Wu, L., Dietrich, U., Mackay, C. R., & Mackay, I. R. (2000) Monoclonal antibody screening of a phage-displayed random peptide library reveals mimotopes of chemokine receptor CCR5: Implications for the tertiary structure of the receptor and for an n-terminal binding site for HIV-1 gp120. European Journal of Immunology, 30(4), pp. 1162-1171.
The chemokine receptor CCR5 contains seven transmembrane-spanning domains. It binds chemokines and acts as co-receptor for macrophage (m)-tropic (or R5) strains of HIV-1. Monoclonal antibodies (mAb) to CCR5, 3A9 and 5C7, were used for biopanning a nonapeptide cysteine (C)-constrained phage-displayed random peptide library to ascertain contact residues and define tertiary structures of possible epitopes on CCR5. Reactivity of antibodies with phagotopes was established by enzyme-linked immunosorbent assay (ELISA). mAb 3A9 identified a phagotope C-HASIYDFGS-C (3A9/1), and 5C7 most frequently identified C-PHWLRDLRV-C (5C7/1). Corresponding peptides were synthesized. Phagotopes and synthetic peptides reacted in ELISA with corresponding antibodies and synthetic peptides inhibited antibody binding to the phagotopes. Reactivity by immunofluorescence of 3A9 with CCR5 was strongly inhibited by the corresponding peptide. Both mAb 3A9 and 5C7 reacted similarly with phagotopes and the corresponding peptide selected by the alternative mAb. The sequences of peptide inserts of phagotopes could be aligned as mimotopes of the sequence of CCR5. For phage 3A9/1, the motif SIYD aligned to residues at the N terminus and FG to residues on the first extracellular loop; for 5C7/1, residues at the N terminus, first extracellular loop, and possibly the third extracellular loop could be aligned and so would contribute to the mimotope. The synthetic peptides corresponding to the isolated phagotopes showed a CD4-dependent reactivity with gp120 of a primary, m-tropic HIV-1 isolate. Thus reactivity of antibodies raised to CCR5 against phage-displayed peptides defined mimotopes that reflect binding sites for these antibodies and reveal a part of the gp120 binding sites on CCR5.
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|Item Type:||Journal Article|
|Keywords:||Chemokine receptor CCR5, Gp120, HIV-1, Mimotope, Phage-displayed peptide library, CD4 antigen, glycoprotein gp 120, monoclonal antibody, peptide, synthetic peptide, amino terminal sequence, animal cell, antigen antibody reaction, article, binding site, controlled study, enzyme linked immunosorbent assay, Human immunodeficiency virus 1, immunofluorescence, mouse, nonhuman, peptide synthesis, phage display, priority journal, protein domain, protein tertiary structure, Amino Acid Sequence, Animals, Antibodies, Monoclonal, Antibody Affinity, Antibody Specificity, B-Lymphocytes, Binding, Competitive, Cloning, Molecular, Enzyme-Linked Immunosorbent Assay, Epitopes, HIV Envelope Protein gp120, Mice, Models, Molecular, Molecular Mimicry, Peptide Library, Peptides, Protein Binding, Protein Structure, Tertiary, Receptors, CCR5, Sequence Analysis, Protein, Tumor Cells, Cultured|
|Copyright Owner:||Copyright 2000 WILEY-VCH Verlag GmbH, Weinheim, Fed. Rep. of Germany|
|Deposited On:||30 Sep 2015 05:51|
|Last Modified:||30 Sep 2015 05:51|
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