Abstract
Gp130 cytokine receptor is involved in the formation of multimeric functional receptors for interleukin-6 (IL-6), IL-11, leukemia inhibitory factor (LIF), oncostatin M (OSM), ciliary neurotrophic factor, and cardiotrophin-1. Cloning of the epitope recognized by an OSM-neutralizing anti-gp130 monoclonal antibody identified a portion of gp130 receptor localized in the EF loop of the cytokine binding domain. Site-directed mutagenesis of the corresponding region was carried out by alanine substitution of residues 186-198. To generate type 1 or type 2 OSM receptors, gp130 mutants were expressed together with either LIF receptor beta or OSM receptor beta. When positions Val-189/Tyr-190 and Phe-191/Val-192 were alanine-substituted, Scatchard analyses indicated a complete abrogation of OSM binding to both type receptors. Interestingly, binding of LIF to type 1 receptor was not affected, corroborating the notion that in this case gp130 mostly behaves as a converter protein rather than a binding receptor. The present study demonstrates that positions 189-192 of gp130 cytokine binding domain are essential for OSM binding to both gp130/LIF receptor beta and gp130/OSM receptor beta heterocomplexes.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Alanine / chemistry
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Amino Acid Sequence
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Animals
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Antibodies, Monoclonal / metabolism
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Antigens, CD / chemistry*
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Antigens, CD / metabolism*
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Binding Sites
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COS Cells
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Cytokine Receptor gp130
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Cytokines / metabolism*
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Dose-Response Relationship, Drug
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Enzyme-Linked Immunosorbent Assay
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Epitopes / metabolism
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Flow Cytometry
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Growth Inhibitors*
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Humans
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Interleukin-6*
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Kinetics
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Leukemia Inhibitory Factor
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Leukemia Inhibitory Factor Receptor alpha Subunit
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Lymphokines*
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Membrane Glycoproteins / chemistry*
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Membrane Glycoproteins / metabolism*
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Models, Molecular
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Molecular Sequence Data
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Mutagenesis, Site-Directed
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Oncostatin M
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Peptide Library
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Peptides / metabolism*
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Protein Binding
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Protein Structure, Secondary
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Receptors, Cytokine / metabolism
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Receptors, OSM-LIF
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Recombinant Proteins / metabolism
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Sequence Homology, Amino Acid
Substances
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Antibodies, Monoclonal
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Antigens, CD
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Cytokines
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Epitopes
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Growth Inhibitors
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IL6ST protein, human
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Interleukin-6
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LIF protein, human
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LIFR protein, human
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Leukemia Inhibitory Factor
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Leukemia Inhibitory Factor Receptor alpha Subunit
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Lymphokines
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Membrane Glycoproteins
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OSM protein, human
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Peptide Library
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Peptides
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Receptors, Cytokine
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Receptors, OSM-LIF
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Recombinant Proteins
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Oncostatin M
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Cytokine Receptor gp130
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Alanine