A role for the immunoglobulin-like domain of the human IL-6 receptor. Intracellular protein transport and shedding

Eur J Biochem. 1999 Jul;263(2):438-46. doi: 10.1046/j.1432-1327.1999.00511.x.

Abstract

Interleukin (IL)-6, IL-11 and cililary neurotrophic factor (CNTF) belong to the same family of hematopoietic and neurotrophic cytokines. Their receptor complexes contain a cytokine-binding alpha receptor and the common glycoprotein (gp)130 subunit for signal transduction. The extracellular parts of the alpha-receptor subunits consist of a membrane-proximal cytokine-binding domain and an N-terminal immunoglobulin (Ig)-like domain with unknown function. We examined the role of the Ig-like domain of IL-6R by constructing deletion mutants lacking the Ig domain (IL-6RDeltaIg and soluble IL-6RDeltaIg). IL-6RDeltaIg was shed as effectively as wild-type IL-6R from transfected COS-7 cells upon 4beta-phorbol 12-myristate 13-acetate (PMA) treatment, whereas nonstimulated shedding of IL-6RDeltaIg was not observed. The shed sIL-6RDeltaIg from PMA-treated cells, as well as the transmembrane IL-6RDeltaIg, had the same biological activity as wild-type sIL-6R, as measured by the induction of haptoglobin secretion in HepG2-IL-6 cells and IL-6-dependent proliferation of IL-6RDeltaIg transfected BAF/gp130 cells. In COS-7 cells transfected with IL-6RDeltaIg or soluble IL-6RDeltaIg cDNA, transport of the deletion mutants through the secretory pathway appeared to be delayed because a sizeable proportion of the mutants was detected as an endo-beta-N-acetylglucosaminidase-sensitive intermediate, suggesting that transport and processing of the DeltaIg mutants on the secretory pathway were impaired. These experiments suggest that the Ig-like domain of the IL-6R is important for intracellular transport of IL-6R through the secretory pathway. Furthermore, the Ig-like domain is necessary for noninduced shedding of the IL-6R, whereas it has no function in PKC-dependent shedding of the IL-6R.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • COS Cells
  • Cell Line
  • Dose-Response Relationship, Drug
  • Flow Cytometry
  • Glycosylation
  • Humans
  • Interleukin-6 / pharmacology
  • Lysosomal Membrane Proteins
  • Membrane Glycoproteins / metabolism
  • Mice
  • Models, Biological
  • Receptors, Interleukin-6 / chemistry*
  • Recombinant Proteins
  • Signal Transduction
  • Tetradecanoylphorbol Acetate / pharmacology
  • Time Factors
  • Transfection

Substances

  • Interleukin-6
  • LAMP1 protein, human
  • Lamp1 protein, mouse
  • Lysosomal Membrane Proteins
  • Membrane Glycoproteins
  • Receptors, Interleukin-6
  • Recombinant Proteins
  • Tetradecanoylphorbol Acetate