Functional expression of soluble human interleukin-11 (IL-11) receptor alpha and stoichiometry of in vitro IL-11 receptor complexes with gp130.
The interleukin-6 (IL-6) family of cytokines activates signaling through the formation of either gp130 homodimers, as for IL-6, or gp130-leukemia inhibitory factor receptor (LIFR) heterodimers as for ciliary neurotrophic factor (CNTF), leukemia inhibitory factor, oncostatinM, and cardiotrophin-1. Recent in vitro studies with IL-6 and CNTF have demonstrated that higher order hexameric receptor complexes are assembled in which signaling chain dimerization is accompanied by the dimerization of both the cytokine molecule and its specific receptor alpha subunits (IL-6Ralpha or CNTFRalpha, respectively). IL-11 is a member of the IL-6 family and known to require gp130 but not LIFR for signaling. In this study we investigate the functional and biochemical composition of the IL-11 receptor complex. The human IL-11 receptor alpha-chain was cloned from a human bone marrow cDNA library. IL-11Ralpha was shown to confer IL-11 responsiveness to human hepatoma cells either by cDNA transfection or by adding a soluble form of the receptor (sIL11Ralpha) expressed in the baculovirus system to the culture medium. In vitro immunoprecipitation experiments showed that sIL11Ralpha specifically binds IL-11 and that binding is enhanced by gp130. Similarly to IL-6 and CNTF, gp130 is able to induce dimerization of the IL-11.IL-11Ralpha subcomplex, the result of which is the formation of a pentameric receptor complex. However, in contrast to the other two cytokines, IL-11 was unable to induce either gp130 homodimerization or gp130/LIFR heterodimerization. These results strongly suggest that an as yet unidentified receptor beta-chain is involved in IL-11 signaling.
Neddermann
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Istituto di Ricerche di Biologia Molecolare "P. Angeletti" (IRBM), Department of Genetics, 00040 Pomezia (Roma), Italy. Neddermann@IRBM.it
Graziani
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R
Ciliberto
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Paonessa
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D000818:Animals; D015703:Antigens, CD / Q000737:chemistry*; D003001:Cloning, Molecular; D050822:Cytokine Receptor gp130; D018076:DNA, Complementary / Q000235:genetics; D006801:Humans; D017370:Interleukin-11 / Q000378:metabolism; D053703:Interleukin-11 Receptor alpha Subunit; D008114:Liver Neoplasms, Experimental; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins / Q000737:chemistry*; D011485:Protein Binding; D018123:Receptors, Interleukin / Q000737:chemistry*; D053702:Receptors, Interleukin-11; D015398:Signal Transduction; D012995:Solubility; D014407:Tumor Cells, Cultured
D015703:Antigens, CD; D018076:DNA, Complementary; C508428:IL11RA protein, human; C494199:IL6ST protein, human; D017370:Interleukin-11; D053703:Interleukin-11 Receptor alpha Subunit; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins; D018123:Receptors, Interleukin; D053702:Receptors, Interleukin-11; D050822:Cytokine Receptor gp130