Identification and characterization of receptor for cytokine-induced neutrophil chemoattractant-3 on rat neutrophils

Biochem Biophys Res Commun. 1997 Mar 17;232(2):562-7. doi: 10.1006/bbrc.1997.6136.

Abstract

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.

MeSH terms

  • Animals
  • Chemokine CXCL2
  • Chemokines, CXC*
  • Chemotactic Factors / genetics
  • Chemotactic Factors / metabolism*
  • Cross-Linking Reagents
  • Growth Substances / genetics
  • Growth Substances / metabolism*
  • Intercellular Signaling Peptides and Proteins*
  • Mutagenesis, Site-Directed
  • Neutrophils / metabolism*
  • Radioligand Assay
  • Rats
  • Receptors, Cytokine / blood*
  • Receptors, Cytokine / chemistry*
  • Receptors, Cytokine / genetics
  • Tyrosine / metabolism

Substances

  • Chemokine CXCL2
  • Chemokines, CXC
  • Chemotactic Factors
  • Cross-Linking Reagents
  • Cxcl2 protein, rat
  • Growth Substances
  • Intercellular Signaling Peptides and Proteins
  • Receptors, Cytokine
  • Tyrosine