gp130 dimerization in the absence of ligand

preformed cytokine receptor complexes

Stephanie Tenhumberg, Björn Schuster, Lixin Zhu, Marina Kovaleva, Jürgen Scheller, Karl-Josef Kallen, Stefan Rose-John, Marina Kovaleva

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

It is established that cytokine receptors signal after ligand binding as homo- or hetero-dimers in heteromeric complexes, but it is unclear, when dimerization occurs. To investigate gp130 dimerization, we performed co-precipitation experiments with the endogenous cytokine receptors gp130 and leukemia inhibitory factor receptor (LIF-R) and with gp130 variants carrying two different C-terminal peptide tags. Furthermore, fluorescence resonance energy transfer (FRET) was employed to detect dimerization of two fluorescent-tagged gp130 variants. Confocal laser scanning microscopy was used for FRET detection in live cells. gp130 and LIF-R could be coprecipitated in the absence of ligand. The interaction, however, was intensified by the addition of LIF. Similar results were obtained with the gp130 variants and confirmed by FRET analysis in live cells. The present study clearly demonstrates the existence of preformed but inactive gp130/LIF-R hetero- and gp130/gp130 homo-dimers. The addition of ligand enhanced the respective dimer formation and was required for signal transduction.
Original languageEnglish
Pages (from-to)649-57
Number of pages9
JournalBiochemical and Biophysical Research Communications
Volume346
Issue number3
DOIs
Publication statusPublished - 4 Aug 2006

Fingerprint

OSM-LIF Receptors
Fluorescence Resonance Energy Transfer
Cytokine Receptors
Dimerization
Dimers
Ligands
Cytokine Receptor gp130
Confocal Microscopy
Signal transduction
Signal Transduction
Coprecipitation
Microscopic examination
Peptides
Scanning
Lasers
Experiments

Keywords

  • Animals
  • Cell Line
  • Cercopithecus aethiops
  • Cytokine Receptor gp130
  • Dimerization
  • Fluorescence Resonance Energy Transfer
  • Humans
  • Leukemia Inhibitory Factor Receptor alpha Subunit
  • Ligands
  • Protein Binding
  • Receptors, Cytokine
  • Receptors, OSM-LIF

Cite this

Tenhumberg, S., Schuster, B., Zhu, L., Kovaleva, M., Scheller, J., Kallen, K-J., ... Kovaleva, M. (2006). gp130 dimerization in the absence of ligand: preformed cytokine receptor complexes. Biochemical and Biophysical Research Communications, 346(3), 649-57. https://doi.org/10.1016/j.bbrc.2006.05.173

gp130 dimerization in the absence of ligand : preformed cytokine receptor complexes. / Tenhumberg, Stephanie; Schuster, Björn; Zhu, Lixin; Kovaleva, Marina; Scheller, Jürgen; Kallen, Karl-Josef; Rose-John, Stefan; Kovaleva, Marina.

In: Biochemical and Biophysical Research Communications, Vol. 346, No. 3, 04.08.2006, p. 649-57.

Research output: Contribution to journalArticle

Tenhumberg, S, Schuster, B, Zhu, L, Kovaleva, M, Scheller, J, Kallen, K-J, Rose-John, S & Kovaleva, M 2006, 'gp130 dimerization in the absence of ligand: preformed cytokine receptor complexes', Biochemical and Biophysical Research Communications, vol. 346, no. 3, pp. 649-57. https://doi.org/10.1016/j.bbrc.2006.05.173
Tenhumberg, Stephanie ; Schuster, Björn ; Zhu, Lixin ; Kovaleva, Marina ; Scheller, Jürgen ; Kallen, Karl-Josef ; Rose-John, Stefan ; Kovaleva, Marina. / gp130 dimerization in the absence of ligand : preformed cytokine receptor complexes. In: Biochemical and Biophysical Research Communications. 2006 ; Vol. 346, No. 3. pp. 649-57.
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AB - It is established that cytokine receptors signal after ligand binding as homo- or hetero-dimers in heteromeric complexes, but it is unclear, when dimerization occurs. To investigate gp130 dimerization, we performed co-precipitation experiments with the endogenous cytokine receptors gp130 and leukemia inhibitory factor receptor (LIF-R) and with gp130 variants carrying two different C-terminal peptide tags. Furthermore, fluorescence resonance energy transfer (FRET) was employed to detect dimerization of two fluorescent-tagged gp130 variants. Confocal laser scanning microscopy was used for FRET detection in live cells. gp130 and LIF-R could be coprecipitated in the absence of ligand. The interaction, however, was intensified by the addition of LIF. Similar results were obtained with the gp130 variants and confirmed by FRET analysis in live cells. The present study clearly demonstrates the existence of preformed but inactive gp130/LIF-R hetero- and gp130/gp130 homo-dimers. The addition of ligand enhanced the respective dimer formation and was required for signal transduction.

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