title: High-affinity binding of bioactive glycosylation-inhibiting factor to antigen-primed T cells and natural killer cells.
abstract: High-affinity binding was demonstrated between suppressor-T-cell-derived bioactive glycosylation-inhibiting factor (GIF) and helper T hybridomas and natural killer cell line cells. Inactive GIF present in cytosol of suppressor T cells and Escherichia coli-derived recombinant human GIF (rhGIF) failed to bind to these cells. However, affinity of rhGIF for the target cells was generated by replacement of Cys-57 in the sequence with Ala or of Asn-106 with Ser or binding of 5-thio-2-nitrobenzoic acid to Cys-60 in the molecule. Such mutations and the chemical modification of rhGIF synergistically increased the affinity of GIF molecules for the target cells. The results indicated that receptors on the target cells recognize conformational structures of bioactive GIF. Equilibrium dissociation constant (Kd) of the specific binding between bioactive rGIF derivatives and high-affinity receptors was 10-100 pM. Receptors for bioactive GIF derivatives were detected on Th1 and Th2 T helper clones and natural killer NK1.1(+) cells in normal spleen but not on naive T or B cells. Neither the inactive rGIF nor bioactive rGIF derivatives bound to macrophage and monocyte lines or induced macrophages for tumor necrosis factor alpha production.
authors.affiliation: Division of Immunobiology, La Jolla Institute for Allergy and Immunology, San Diego, CA 92121, USA.
authors.forename: K
authors.initials: K
authors.lastname: Sugie
authors.forename: T
authors.initials: T
authors.lastname: Nakano
authors.forename: T
authors.initials: T
authors.lastname: Tomura
authors.forename: K
authors.initials: K
authors.lastname: Takakura
authors.forename: T
authors.initials: T
authors.lastname: Mikayama
authors.forename: K
authors.initials: K
authors.lastname: Ishizaka
mesh_terms: D000409:Alanine; D000818:Animals; D001216:Asparagine; D001665:Binding Sites; D002460:Cell Line; D003545:Cysteine; D004926:Escherichia coli; D006801:Humans; D006825:Hybridomas; D007694:Killer Cells, Natural / Q000276:immunology* / Q000378:metabolism; D007700:Kinetics; D008213:Lymphocyte Activation; D008222:Lymphokines / Q000378:metabolism* / Q000494:pharmacology; D008264:Macrophages; D051379:Mice; D008807:Mice, Inbred BALB C; D016297:Mutagenesis, Site-Directed; D017354:Point Mutation; D029584:Prostatic Secretory Proteins; D011994:Recombinant Proteins / Q000378:metabolism; D012694:Serine; D013491:Suppressor Factors, Immunologic / Q000378:metabolism / Q000494:pharmacology; D050378:T-Lymphocytes, Regulatory / Q000276:immunology* / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
chemical_list: D008222:Lymphokines; D029584:Prostatic Secretory Proteins; D011994:Recombinant Proteins; D013491:Suppressor Factors, Immunologic; D014409:Tumor Necrosis Factor-alpha; C043076:beta-microseminoprotein; C013059:immunoglobulin-binding factors; D012694:Serine; D001216:Asparagine; D003545:Cysteine; D000409:Alanine