N-terminal domain of eotaxin-3 is important for activation of CC chemokine receptor 3.
Eotaxin-3 belongs to the CC chemokine family, and specifically recognizes CC chemokine receptor (CCR) 3 that is expressed on eosinophils, basophils and helper T type 2 cells. The three-dimensional structure of eotaxin-3 determined by nuclear magnetic resonance has revealed that the N-terminal nine residues preceding the first cysteine comprise an unstructured domain, which is also observed in other chemokine molecules. In order to determine the function of the N-terminal domain of eotaxin-3, we constructed various N-terminal-deletion mutants, and then examined their binding and chemotactic activities toward eosinophils in vitro. Competitive binding studies showed that the binding affinity of truncated mutant toward CCR3 was almost the same as that of wild-type eotaxin-3 even though the N-terminal truncation involved the first through to the ninth residues. In contrast, the chemotactic activity gradually decreased with extension of the N-terminal deletion, and when the deletion extended to the eighth residue, the activity was not detected at all. Thus, the N-terminal nine residues are not critical for binding but the N-terminal eight residues are essential for activation of CCR3. The truncated eotaxin-3 proteins lacking the N-terminal eight or nine residues inhibited the chemotactic activity of chemokines that recognize CCR3. The truncated mutants can possibly be used for anti-allergic and anti-HIV-1 therapy.
Shinkai
Akeo
A
Tokyo Research Laboratories, Kyowa Hakko Kogyo Co, Ltd, 3-6-6 Asahi-machi, Machida-shi, Tokyo 194-8533, Japan. ashinkai@sp8sun.spring8.or.jp
Komuta-Kunitomo
Mayumi
M
Sato-Nakamura
Naoko
N
Anazawa
Hideharu
H
D002465:Cell Movement / Q000235:genetics / Q000502:physiology; D000075906:Chemokine CCL26; D019742:Chemokines, CC / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004804:Eosinophils / Q000378:metabolism; D006801:Humans; D017434:Protein Structure, Tertiary / Q000502:physiology; D054397:Receptors, CCR3; D019707:Receptors, Chemokine / Q000378:metabolism*; D017384:Sequence Deletion / Q000502:physiology; D013329:Structure-Activity Relationship
C120429:CCL26 protein, human; C515350:CCR3 protein, human; D000075906:Chemokine CCL26; D019742:Chemokines, CC; D054397:Receptors, CCR3; D019707:Receptors, Chemokine