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Receptors for human gamma interferon: binding and crosslinking of 125I-labeled recombinant human gamma interferon to receptors on WISH cells. 1984

F H Sarkar, and S L Gupta

Purified recombinant human gamma interferon (HuIFN-gamma), labeled with 125I (125I-HuIFN-gamma), was used to study receptors for HuIFN-gamma on human WISH cells. 125I-HuIFN-gamma was bound to WISH cells, and this binding was displaced by unlabeled HuIFN-gamma but not by unlabeled recombinant HuIFN-alpha 2 or [Ser17]HuIFN-beta (HuIFN-beta with serine substituted for cysteine at position 17), indicating the presence of specific binding sites for HuIFN-gamma. The cell-bound 125I-HuIFN-gamma was crosslinked with disuccinimidyl suberate or ethylene glycol bis(succinimidyl succinate), which yielded a complex of Mr approximately 105,000 +/- 5000 as analyzed by NaDodSO4/PAGE. The formation of this complex was prevented by preincubation of cells with unlabeled HuIFN-gamma but not with HuIFN-alpha 2 or [Ser17]HuIFN-beta, indicating that HuIFN-gamma binds to a specific receptor molecule and that HuIFN-alpha 2 or HuIFN-beta do not interact with this receptor. Experiments were carried out with 125I-labeled recombinant [Ser17]HuIFN-beta (125I-[Ser17]HuIFN-beta) to verify this conclusion. Binding and crosslinking of 125I-[Ser17]HuIFN-beta to human WISH cells and Daudi cells yielded a complex of Mr approximately 150,000 similar to that obtained with 125I-HuIFN-alpha 2 as described earlier. The formation of this Mr 150,000 complex with 125I-[Ser17]HuIFN-beta was displaced by unlabeled [Ser17]HuIFN-beta and by HuIFN-alpha 2 but not by HuIFN-gamma, indicating that [Ser17]HuIFN-beta binds to the same receptor as does HuIFN-alpha 2, identified earlier, and that HuIFN-gamma does not compete with 125I-[Ser17]HuIFN-beta for this receptor. We conclude that HuIFN-gamma interacts with specific receptors that are distinctly different from the receptors recognized by HuIFN-alpha and HuIFN-beta.

UI MeSH Term Description Entries
D007371 Interferon-gamma The major interferon produced by mitogenically or antigenically stimulated LYMPHOCYTES. It is structurally different from TYPE I INTERFERON and its major activity is immunoregulation. It has been implicated in the expression of CLASS II HISTOCOMPATIBILITY ANTIGENS in cells that do not normally produce them, leading to AUTOIMMUNE DISEASES. Interferon Type II,Interferon, Immune,gamma-Interferon,Interferon, gamma,Type II Interferon,Immune Interferon,Interferon, Type II
D007700 Kinetics The rate dynamics in chemical or physical systems.
D008970 Molecular Weight The sum of the weight of all the atoms in a molecule. Molecular Weights,Weight, Molecular,Weights, Molecular
D011247 Pregnancy The status during which female mammals carry their developing young (EMBRYOS or FETUSES) in utero before birth, beginning from FERTILIZATION to BIRTH. Gestation,Pregnancies
D011956 Receptors, Cell Surface Cell surface proteins that bind signalling molecules external to the cell with high affinity and convert this extracellular event into one or more intracellular signals that alter the behavior of the target cell (From Alberts, Molecular Biology of the Cell, 2nd ed, pp693-5). Cell surface receptors, unlike enzymes, do not chemically alter their ligands. Cell Surface Receptor,Cell Surface Receptors,Hormone Receptors, Cell Surface,Receptors, Endogenous Substances,Cell Surface Hormone Receptors,Endogenous Substances Receptors,Receptor, Cell Surface,Surface Receptor, Cell
D002460 Cell Line Established cell cultures that have the potential to propagate indefinitely. Cell Lines,Line, Cell,Lines, Cell
D005260 Female Females
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000650 Amnion The innermost membranous sac that surrounds and protects the developing embryo which is bathed in the AMNIOTIC FLUID. Amnion cells are secretory EPITHELIAL CELLS and contribute to the amniotic fluid. Amniotic Membrane,Amnions,Amniotic Membranes,Membrane, Amniotic,Membranes, Amniotic
D001667 Binding, Competitive The interaction of two or more substrates or ligands with the same binding site. The displacement of one by the other is used in quantitative and selective affinity measurements. Competitive Binding

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