Identification and characterization of receptor for cytokine-induced neutrophil chemoattractant-3 on rat neutrophils.
Cytokine-induced neutrophil chemoattractants (CINCs), members of alpha-chemokines, are considered to be major neutrophil chemotactic factors in rats. Recombinant CINC-3/rat macrophage inflammatory protein-2 (MIP-2) having an additional tyrosine residue at the carboxyl terminus (CINC-3-Tyr) was constructed, purified, radiolabeled with 125I, and used for binding studies. The specific binding of 125I-labeled CINC-3-Tyr (125I-CINC-3-Tyr) to rat neutrophils reached a plateau after approximately 60 min at 4 degrees C. This binding of 125I-CINC-3-Tyr could be reversed by adding an excess amount of unlabeled CINC-3-Tyr. Scatchard analysis revealed approximately 12,000 binding sites per cell on rat neutrophils with an apparent Kd value of 120 pM. Chemical cross-linking experiments suggested that the rat neutrophil CINC-3 receptor is a mass of approximately 69 kDa. Taken together, our results strongly suggest that rat neutrophils express a high affinity receptor for CINC-3.
Murakami
K
K
Department of Pharmacology, Green Cross Corporation, Osaka, Japan. gcc39616@greencross.co.jp
Shibata
F
F
al-Mokdad
M
M
Nakagawa
H
H
Ueno
A
A
Kondo
T
T
D000818:Animals; D054426:Chemokine CXCL2; D019743:Chemokines, CXC; D002630:Chemotactic Factors / Q000235:genetics / Q000378:metabolism*; D003432:Cross-Linking Reagents; D006133:Growth Substances / Q000235:genetics / Q000378:metabolism*; D036341:Intercellular Signaling Peptides and Proteins; D016297:Mutagenesis, Site-Directed; D009504:Neutrophils / Q000378:metabolism*; D011869:Radioligand Assay; D051381:Rats; D018121:Receptors, Cytokine / Q000097:blood* / Q000737:chemistry* / Q000235:genetics; D014443:Tyrosine / Q000378:metabolism
D054426:Chemokine CXCL2; D019743:Chemokines, CXC; D002630:Chemotactic Factors; D003432:Cross-Linking Reagents; C513296:Cxcl2 protein, rat; D006133:Growth Substances; D036341:Intercellular Signaling Peptides and Proteins; D018121:Receptors, Cytokine; D014443:Tyrosine