The N-Terminus of gp130 is Critical for the Formation of the High-Affinity Interleukin-6 Receptor Complex Academic Article uri icon

abstract

  • Interleukin-6 (IL-6) mediates its activity through binding to two cell-surface receptors. The high-affinity human IL-6 receptor complex consists of two transmembrane anchored subunits: a ligand-specific, low-affinity IL-6 receptor and the high-affinity converter and signal transducing, gp130. Previously, using recombinant forms of human IL-6 and the extracellular ('soluble') domains of the IL-6 receptor (sIL-6R) and gp130 (sgp130), we have shown that the high-affinity IL-6R complex is hexameric, consisting of two molecules each of IL-6, sIL-6R and sgp130 (Ward et al., 1994, J. Biol. Chem. 269: 23286-23289). This paper investigates the role of the N-terminal region of gp130 in the formation of the high-affinity IL-6R complex. Using recombinant sgp130 produced with a FLAG octapeptide epitope (DYKDDDDK) at the N-terminus (sgp130-FLAG), we demonstrate, using biosensor analysis and size-exclusion chromatography, that modification of the N-terminus of sgp130 interferes with the in vitro in solution formation of the stable hexameric IL-6 receptor complex. Rather, sgp130-FLAG interacts with IL-6 and sIL-6R with a much lower affinity and forms a stable lower-order ternary complex. However, this lower-order complex is inconsistent with the solution molecular weight of a trimeric complex, as measured by size-exclusion chromatography. In contrast, N-terminal modification of the sgp130 with the FLAG epitope did not interfere with the binding of leukemia inhibitory factor or oncostatin-M (other cytokines that signal through gp130) to sgp130. These data support our model of the hexameric IL-6 receptor complex, which is biased towards the association of two IL-6.IL-6R.gp130 trimers, and postulates the critical involvement of the N-terminal Ig-like domain of gp130 in tethering the two trimers to form the stable hexamer (Simpson et al., 1997, Prot. Sci. 6: 929-955).

authors

  • Moritz, Robert L
  • Ward, Larry D
  • Tu, Guo-Fen
  • Fabri, Louis J
  • Ji, Hong
  • Yasukawa, Kiyoshi
  • Simpson, Richard J

publication date

  • January 1999

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