Solubilization and characterization of erythropoietin receptor from transplantable mouse erythroblastic leukemic cells

Biochem Biophys Res Commun. 1989 May 15;160(3):1140-8. doi: 10.1016/s0006-291x(89)80122-6.

Abstract

Functional erythropoietin (Ep) receptor (Ep-R) on transplantable mouse leukemic cells was solubilized by 3[(3-cholamidopropyl)dimethyl ammonio]-1-propanesulfate (CHAPS). An assay of the solubilized Ep-R was established: (1) radioiodinated Ep was bound to the solubilized Ep-R, (2) Ep.Ep-R complexes were covalently cross-linked, (3) the cross-linked products were selectively precipitated by polyethylene glycol, (4) the products were separated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and (5) the Ep.Ep-R cross-linked products were measured. Optimal conditions to yield the cross-linked products and to stabilize the solubilized Ep-R were found. The solubilized Ep-R had properties similar to those of Ep-R on the membrane with respect to binding with Ep and the cross-linked products. These results would be exploited to allow purification of Ep-R.

MeSH terms

  • Animals
  • Cell Membrane / analysis
  • Chemical Precipitation
  • Cholic Acids
  • Cross-Linking Reagents
  • Electrophoresis, Polyacrylamide Gel
  • Erythropoietin / metabolism
  • Leukemia, Erythroblastic, Acute / metabolism*
  • Mice
  • Mice, Inbred C3H
  • Neoplasm Transplantation
  • Polyethylene Glycols
  • Receptors, Cell Surface / analysis*
  • Receptors, Cell Surface / metabolism
  • Receptors, Erythropoietin
  • Solubility
  • Succinimides

Substances

  • Cholic Acids
  • Cross-Linking Reagents
  • Receptors, Cell Surface
  • Receptors, Erythropoietin
  • Succinimides
  • Erythropoietin
  • Polyethylene Glycols
  • 3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate
  • disuccinimidyl suberate