title: Cross-linking of human neutrophil surface proteins to iodinated interleukin 8 or neutrophil activating peptide-2 results in at least four separable proteins.
abstract: The human neutrophil activating peptides-1 and -2 (NAP-1/IL-8, NAP-2) are two structurally and functionally related members of the chemokine cytokine family. They are chemoattractants and activators of neutrophils and exert their effects by binding to specific receptors which are expressed on responsive cells. Two closely related IL-8 receptors of neutrophils have been characterized recently by molecular cloning. We show here that NAP-1/IL-8 and NAP-2 can be cross-linked to at least four protein bands from human neutrophil surfaces with apparent molecular masses of 55, 65, 71 and 81 kDa. The two cross-linked proteins with lower masses were associated with high, the two with the higher masses with low affinity binding of NAP-2, NAP-1/IL-8 was bound to all bands with high affinity. NAP-1/IL-8 and NAP-2 could also be cross-linked to form dimers when bound to cells and in solution. Our results show that more than two NAP-1/IL-8 receptors, or more than two forms of the known receptors exist. Alternatively, the four protein bands can be explained by cross-linking of ligand monomers and dimers, respectively, to the known receptors of neutrophils.
authors.affiliation: Sandoz Forschungsinstitut, Vienna, Austria.
authors.forename: J
authors.initials: J
authors.lastname: Besemer
authors.forename: W
authors.initials: W
authors.lastname: Schnitzel
authors.forename: U
authors.initials: U
authors.lastname: Monschein
authors.forename: B
authors.initials: B
authors.lastname: Ryffel
mesh_terms: D003432:Cross-Linking Reagents; D016207:Cytokines / Q000302:isolation & purification*; D006801:Humans; D066298:In Vitro Techniques; D016209:Interleukin-8 / Q000737:chemistry*; D007457:Iodine Radioisotopes; D010455:Peptides / Q000737:chemistry / Q000187:drug effects*; D001620:beta-Thromboglobulin
chemical_list: D003432:Cross-Linking Reagents; D016207:Cytokines; D016209:Interleukin-8; D007457:Iodine Radioisotopes; C061615:PPBP protein, human; D010455:Peptides; D001620:beta-Thromboglobulin; C009862:connective tissue-activating peptide