Analysis of the Leukemia Inhibitory Factor Receptor Functional Domains by Chimeric Receptors and Cytokines

• Published in: Biochemistry (Easton) Vol. 42; no. 18; pp. 5244 - 5252
• Main Authors: Aasland, Dorthe, Schuster, Björn, Grötzinger, Joachim, Rose-John, Stefan, Kallen, Karl-Josef
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 13.05.2003
• Subjects: Animals; Binding Sites; Cercopithecus aethiops; COS Cells; Immunoglobulins - chemistry; Interleukin-6 - metabolism; Ligands; Lymphokines - chemistry; Lymphokines - genetics; Lymphokines - metabolism; Mutation - genetics; Phosphorylation; Phosphotyrosine - metabolism; Receptors, Cytokine - chemistry; Receptors, Cytokine - genetics; Receptors, Cytokine - metabolism; Receptors, Interleukin-6 - chemistry; Receptors, Interleukin-6 - genetics; Receptors, Interleukin-6 - metabolism; Receptors, OSM-LIF; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Sequence Deletion - genetics; Transfection
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi0263311

In contrast to other hematopoietic cytokine receptors, the leukemia inhibitory factor receptor (LIFR) possesses two cytokine binding modules (CBMs). Previous studies suggested that the NH2-terminal CBM and the Ig-like domain of the LIFR are most important for LIF binding and activity. Using the recently engineered designer cytokine IC7, which induces an active heterodimer of the LIFR and gp130 after binding to the IL-6R, and several receptor chimeras of the LIFR and the interleukin-6 receptor (IL-6R) carrying the CBM of the IL-6R in place of the COOH-terminal LIFR CBM, we could assign individual receptor subdomains to individual binding sites of the ligand. The NH2-terminal CBM and the Ig-like domain of the LIFR bind to ligand site III, whereas the COOH-terminal CBM contacts site I. Furthermore, we show that LIFR mutants carrying the IL-6R CBM instead of the COOH-terminal CBM can replace the IL-6R by acting as an α-receptor for IL-6. However, in situations where a signaling competent receptor is bound at IL-6 site I, ligand binding to site III is an absolute requirement for participation of the receptor in a signaling heterodimer with gp130; i.e., a functional receptor complex of IL-6 type cytokines cannot be assembled solely via site I and II as in the growth hormone receptor complex.