Evidence for the ligand-induced conversion from a dimer to a tetramer of the granulocyte colony-stimulating factor receptor

FEBS Lett. 1994 Dec 19;356(2-3):255-60. doi: 10.1016/0014-5793(94)01278-4.

Abstract

An extracellular portion of granulocyte colony-stimulating factor (G-CSF) receptor, which contains an immunoglobulin-like (Ig) domain and cytokine receptor homologous (CRH) region, was secreted into the medium using Trichoplusia ni-Autographa californica nuclear polyhedrosis virus system. The gene product was purified to homogeneity mainly as a dimer (85 kDa) using G-CSF affinity column chromatography and gel filtration HPLC, although the product existed as a monomer (45 kDa) in the medium. Scatchard analyses suggested that only the dimer had high affinity ligand binding (Kd = about 100 pM), which is comparable with the Kd value of the cell surface receptor. The binding of G-CSF to Ig-CRH induced its tetramerization (200-250 kDa). The molecular composition of the tetrameric complex showed a stoichiometry of four ligands bound to four Ig-CRH. These results suggested that the oligomeric mechanism of the G-CSF receptor differs from that reported for growth hormone (GH) receptor, although CD spectrum spectroscopy suggested that the Ig-CRH has a GH receptor-like structure.

MeSH terms

  • Animals
  • Base Sequence
  • Blotting, Western
  • Cell Line
  • DNA Primers
  • Genetic Vectors
  • Insecta
  • Ligands
  • Macromolecular Substances
  • Mice
  • Molecular Sequence Data
  • Nucleopolyhedroviruses
  • Polymerase Chain Reaction
  • Receptors, Cytokine / chemistry
  • Receptors, Granulocyte Colony-Stimulating Factor / chemistry
  • Receptors, Granulocyte Colony-Stimulating Factor / isolation & purification
  • Receptors, Granulocyte Colony-Stimulating Factor / metabolism*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Sequence Homology, Amino Acid
  • Transfection

Substances

  • DNA Primers
  • Ligands
  • Macromolecular Substances
  • Receptors, Cytokine
  • Receptors, Granulocyte Colony-Stimulating Factor
  • Recombinant Proteins