Ligand binding kinetics of a soluble full length murine erythropoietin receptor

Lena S. Avedissian, Indra Poola, Jerry L. Spivak

Research output: Contribution to journalArticle

Abstract

The full-length murine erythropoietin receptor was expressed in Sf9 cells using a baculovirus vector. Erythropoietin receptors in solubilized Sf9 cell lysates bound erythropoietin with high affinity (92 pM) Erythropoietin receptor-125I-labeled erythropoietin association and dissociation kinetics using solubilized Sf9 cell lysates revealed a ka of 0.16 nM-2 min-1 and a kd of 0.00055 min-1 giving an observed KD of 3.45 pM. The erythropoietin receptor was partially purified from Sf9 cell lysates by chromatography on Con A Sepharose. When erythropoietin receptors were crosslinked to 125I-labeled erythropoietin and analyzed by SDS-7.5% PAGE protein complexes of 90 and 125 kDa were observed with receptors in solubilized lysate, and 170 and 190 kDa with the partially purified receptors.

Original languageEnglish (US)
Pages (from-to)62-68
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume216
Issue number1
DOIs
StatePublished - Jan 1 1995

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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