Peptide mimics of monocyte chemoattractant protein-1 (MCP-1) with an antagonistic activity.
In this study, we attempted to analyze the peptide motifs recognized by 24822.111 and F9, monoclonal antibodies (mAbs) that inhibit the chemotactic activity of monocyte chemoattractant protein-1 (MCP-1), a member of the CC subfamily of chemokines. We isolated phage clones from a phage display library and identified six peptide motifs. One of these clones, C27, was strongly and specifically recognized by 24822.111 mAb, while another, G25, was similarly recognized by F9 mAb. Both the C27 motif and the G25 motif contain two cysteines in their sequences and have little homology to the primary amino acid sequence of MCP-1. These clones, however, bound to THP-1 cells, and the binding was competitively inhibited by MCP-1. The clones strongly inhibited the MCP-1-induced chemotaxis of human monocytes. The synthetic and intramolecularly disulfide-linked peptides of C27 and G25 (sC27 and sG25) also inhibited the chemotaxis induced by MCP-1, while their derivatives with serine in place of cysteine did not, suggesting the importance of the loop structure for the inhibition. These results suggest that sC27 and sG25 may mimic the MCP-1-binding domain to the MCP-1 receptor.
Kaji
M
M
Department of Bioengineering, Faculty of Engineering, Kagoshima University, Korimoto, Kagoshima 890-0065, Japan.
Ikari
M
M
Hashiguchi
S
S
Ito
Y
Y
Matsumoto
R
R
Yoshimura
T
T
Kuratsu Ji
Sugimura
K
K
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D001435:Bacteriophages / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D015153:Blotting, Western; D002460:Cell Line; D018932:Chemokine CCL2 / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000276:immunology / Q000494:pharmacology; D002633:Chemotaxis / Q000187:drug effects; D015195:Drug Design; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D005434:Flow Cytometry; D006801:Humans; D018716:Molecular Mimicry; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D054390:Receptors, CCR2; D019707:Receptors, Chemokine / Q000378:metabolism
D000911:Antibodies, Monoclonal; C515334:CCR2 protein, human; D018932:Chemokine CCL2; D000939:Epitopes; D019151:Peptide Library; D010455:Peptides; D054390:Receptors, CCR2; D019707:Receptors, Chemokine