Structural and functional studies on the human interleukin-6 receptor. Binding, cross-linking, internalization, and degradation of interleukin-6 by fibroblasts transfected with human interleukin-6-receptor cDNA

J Biol Chem. 1991 Feb 25;266(6):3841-6.

Abstract

A cDNA coding for the human interleukin-6 receptor (IL-6-R) has been expressed stably in murine NIH/3T3 fibroblasts. Transfected cells exhibited a single class of binding sites for 125I-labeled recombinant human interleukin-6 (125I-rhIL-6) (Kd = 440 pM, 20,000 receptors per cell). Affinity cross-linking of 125I-rhIL-6 to the IL-6-R-expressing NIH/3T3 cells led to the detection of three 125I-rhIL-6-containing protein complexes with molecular masses of 100, 120, and 200 kDa suggesting a complex organization of the IL-6-R in the plasma membrane. IL-6 added to the transfected NIH/3T3 cells exerted growth inhibition. This anti-growth effect was observed by the measurement of cell numbers and ornithine decarboxylase mRNA expression. IL-6-R overexpressing fibroblasts internalized 125I-rhIL-6. Intracellular limited proteolysis of IL-6 could be demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A possible implication of skin fibroblasts in the catabolism of IL-6 is discussed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autoradiography
  • Cross-Linking Reagents
  • DNA / genetics*
  • Electrophoresis, Agar Gel
  • Fibrinogen / analysis
  • Fibroblasts / metabolism
  • Humans
  • Interleukin-6 / metabolism*
  • Ornithine Decarboxylase / analysis
  • Plasmids
  • Precipitin Tests
  • RNA, Messenger / analysis
  • Receptors, Immunologic / genetics
  • Receptors, Immunologic / metabolism*
  • Receptors, Interleukin-6
  • Recombinant Proteins / metabolism
  • Structure-Activity Relationship
  • Transfection

Substances

  • Cross-Linking Reagents
  • Interleukin-6
  • RNA, Messenger
  • Receptors, Immunologic
  • Receptors, Interleukin-6
  • Recombinant Proteins
  • Fibrinogen
  • DNA
  • Ornithine Decarboxylase