Identification of oligopeptide sequences which inhibit migration induced by a wide range of chemokines.
We have identified an amino acid sequence, termed peptide 3, corresponding to amino acids 51-62 of the mature human monocyte chemoattractant protein-1 (MCP-1), which inhibits human mononuclear-cell and THP-1-cell migration induced by a wide range of chemokines. For example, peptide 3 inhibited MCP-1-induced THP-1 migration in a transwell assay with an ED50 of approx. 8 microM. Peptide 3 binds directly to THP-1 cells with an association constant of approx. 10 microM, and is therefore likely to be a direct receptor antagonist for CC and CXC chemokine receptors. By performing a structure-function analysis of this peptide, we have identified a sequence variant that shows an approx. 3-4-fold greater potency as an inhibitor of chemokine-induced migration [Leu4Ile11 peptide 3 (1-12)]. Furthermore, unlike peptide 3, which binds to the Duffy antigen receptor for chemokines on human erythrocytes with a similar affinity to the specific chemokine receptors on THP-1 cells, the Leu4Ile11 peptide 3 (1-12) sequence variant shows at least 20-fold greater selectivity for the specific receptors. Derivatives of Leu4Ile11 peptide 3 (1-12) are therefore the best candidates among the molecules we have investigated for use as a chemokine inhibitor in vivo.
Reckless
J
J
Department of Medicine, University of Cambridge, Addenbrookes Hospital, Box 157, Hills Road, Cambridge CB2 2QQ, UK. jr219@mole.bio.cam.ac.uk
Grainger
D J
DJ
D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan; D002352:Carrier Proteins / Q000378:metabolism; D002478:Cells, Cultured; D018932:Chemokine CCL2 / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D018925:Chemokines / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D002634:Chemotaxis, Leukocyte / Q000187:drug effects*; D017124:Conserved Sequence / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D004912:Erythrocytes / Q000166:cytology / Q000378:metabolism; D005260:Female; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D010446:Peptide Fragments / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D015800:Protozoan Proteins; D011956:Receptors, Cell Surface / Q000378:metabolism; D019707:Receptors, Chemokine / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000502:physiology; D016415:Sequence Alignment; D013329:Structure-Activity Relationship; D016212:Transforming Growth Factor beta / Q000494:pharmacology
D000953:Antigens, Protozoan; D002352:Carrier Proteins; D018932:Chemokine CCL2; D018925:Chemokines; C066001:Duffy antigen binding protein, Plasmodium; D009842:Oligopeptides; D010446:Peptide Fragments; D015800:Protozoan Proteins; D011956:Receptors, Cell Surface; D019707:Receptors, Chemokine; D016212:Transforming Growth Factor beta