Recruitment of a heparan sulfate subunit to the interleukin-1 receptor complex. Regulation by fibronectin attachment

J Biol Chem. 1999 Jul 16;274(29):20103-9. doi: 10.1074/jbc.274.29.20103.

Abstract

In this study, we identified an adhesion-regulated subunit of the interleukin-1 (IL-1) receptor complex. Transfection of fibroblasts with an IL-1 receptor-EGFP construct showed that the fusion protein was located at focal adhesions in cells attaching to fibronectin. Fibronectin attachment caused enhancement in endogenous IL-1 type I receptor levels from on average 2500 to 4300 receptors/cell. In addition, matrix attachment resulted in a decrease in binding affinity (Ka) from 1.0 x 10(9) (M-1) to 5.6 x 10(8) (M-1), due to a 2-fold reduction in association rate constant. The adhesion-mediated effects were reversed by soluble heparin. Cross-linking experiments showed that in cells attached to fibronectin, 50-70% of the radiolabeled IL-1 was associated with a heparinase sensitive, high molecular mass component of about 300 kDa, with a core protein of 80-90 kDa. Formation of the complex was dependent on cell interaction with the heparin binding region in fibronectin and required IL-1/type I IL-1 receptor binding. This report demonstrates the recruitment of a heparan sulfate to the IL-1 receptor complex, following attachment to fibronectin, which correlates with alterations in receptor function. The data suggest that the heparan sulfate constitutes an attachment regulated component of the IL-1 receptor complex with the role of mediating matrix regulation of IL-1 responses.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Cells, Cultured
  • Fibroblasts / metabolism
  • Fibronectins / metabolism*
  • Green Fluorescent Proteins
  • Heparitin Sulfate / chemistry
  • Heparitin Sulfate / metabolism*
  • Humans
  • Interleukin-1 / metabolism
  • Iodine Radioisotopes
  • Luminescent Proteins / metabolism
  • Protein Binding
  • Radioligand Assay
  • Receptors, Interleukin-1 / metabolism*
  • Recombinant Fusion Proteins / metabolism

Substances

  • Fibronectins
  • Interleukin-1
  • Iodine Radioisotopes
  • Luminescent Proteins
  • Receptors, Interleukin-1
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins
  • Heparitin Sulfate