Peptides from the amino-terminus of RANTES cause chemotaxis of human T-lymphocytes.
RANTES is a potent chemotactic and activating agent for a variety of leukocytes including T-lymphocytes. To identify the region of the molecule responsible for this activity, we have made overlapping 10 amino acid peptides scanning the protein. A micro-Boyden chamber chemotaxis assay showed the most efficacious peptides came from at the N-terminus. EC50 values of 8 nM (+/- 2.2 nM), 3.7 nM (+/- 2.0 nM) and 3.1 nM (+/- 2.0 nM) were calculated from dose response curves for the peptides (1-10), (3-12), and (5-14). Control peptides from other regions are not active. In THP-1 cells, none of the peptides give a Ca2+ response. The active peptides are shown to be principally chemotactic rather than chemokinetic by a checker board analysis. The results imply that the principal region of RANTES responsible for chemotaxis is located in the amino terminus.
Wells
T N
TN
Glaxo Institute for Molecular Biology, Geneva, Switzerland.
Guye-Coulin
F
F
Bacon
K B
KB
D000595:Amino Acid Sequence; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D018946:Chemokine CCL5; D002634:Chemotaxis, Leukocyte / Q000187:drug effects*; D006801:Humans; D008222:Lymphokines / Q000494:pharmacology*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D013601:T-Lymphocytes / Q000187:drug effects / Q000502:physiology*
D018946:Chemokine CCL5; D008222:Lymphokines; D010446:Peptide Fragments; D002118:Calcium