Two different epitopes of the signal transducer gp130 sequentially cooperate on IL-6-induced receptor activation.
Cytokines are key mediators for the regulation of hemopoiesis and the coordination of immune responses. They exert their various functions through activation of specific cell surface receptors, thereby initiating intracellular signal transduction cascades which lead to defined cellular responses. As the common signal-transducing receptor subunit of at least seven different cytokines, gp130 is an important member of the family of hemopoietic cytokine receptors which are characterized by the presence of at least one cytokine-binding module. Mutants of gp130 that either lack the Ig-like domain D1 (DeltaD1) or contain a distinct mutation (F191E) within the cytokine-binding module have been shown to be severely impaired with respect to IL-6 induced signal transduction. After cotransfection of COS-7 cells with a combination of both inactive gp130 mutants, signal transduction in response to IL-6 is restored. Whereas cells transfected with DeltaD1 do not bind IL-6/sIL-6R complexes, cells transfected with the F191E mutant bind IL-6/sIL-6R with low affinity. Combination of DeltaD1 and F191E, however, leads to high-affinity ligand binding. These data suggest that two different gp130 epitopes, one on each receptor chain, sequentially cooperate in asymmetrical binding of IL-6/IL-6R in a tetrameric signaling complex. On the basis of our data, a model for the mechanism of IL-6-induced gp130 activation is proposed.
Pflanz
S
S
Institut für Biochemie, Rheinisch-Westfälische Technische Hochschule Aachen, Aachen, Germany.
Kurth
I
I
Grötzinger
J
J
Heinrich
P C
PC
Müller-Newen
G
G
D020816:Amino Acid Motifs / Q000235:genetics / Q000276:immunology; D000818:Animals; D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D019556:COS Cells; D050822:Cytokine Receptor gp130; D019281:Dimerization; D000939:Epitopes / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D005822:Genetic Vectors / Q000276:immunology; D018698:Glutamic Acid / Q000235:genetics; D006801:Humans; D007136:Immunoglobulins / Q000235:genetics / Q000378:metabolism; D015850:Interleukin-6 / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D008024:Ligands; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D016297:Mutagenesis, Site-Directed; D010649:Phenylalanine / Q000235:genetics; D011485:Protein Binding / Q000235:genetics / Q000276:immunology; D017434:Protein Structure, Tertiary / Q000235:genetics; D019947:Receptors, Interleukin-6 / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion / Q000276:immunology; D015398:Signal Transduction / Q000235:genetics / Q000276:immunology*; D012995:Solubility; D014162:Transfection
D015703:Antigens, CD; D000939:Epitopes; C494199:IL6ST protein, human; D007136:Immunoglobulins; D015850:Interleukin-6; D008024:Ligands; D008562:Membrane Glycoproteins; D019947:Receptors, Interleukin-6; D050822:Cytokine Receptor gp130; D018698:Glutamic Acid; D010649:Phenylalanine