Expression cloning of the murine erythropoietin receptor

doi:10.1016/0092-8674(89)90965-3

Cell

Volume 57, Issue 2, 21 April 1989, Pages 277-285

https://doi.org/10.1016/0092-8674(89)90965-3 Get rights and content

Abstract

Two independent cDNA clones encoding the erythropoietin receptor (EPO-R) were isolated from a pXM expression library made from uninduced murine erythroleukemia (MEL) cells. The clones were identified by screening COS cell transfectants for binding and uptake of radioiodinated recombinant human erythropoietin. As inferred from the cDNA sequence, the murine erythropoietin receptor is a 507 amino acid polypeptide with a single membrane-spanning domain. It shows no similarities to known proteins or nucleic acid sequences in the data bases. Although the MEL cell EPO-R has a single affinity with a dissociation constant of approximately 240 pM, the EPO-R cDNA, expressed in COS cells, generates both a high-affinity (30 pM) and a low-affinity (210 pM) receptor.

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I was convinced that this insane technology would eventually work, and my role in the project became mainly that of cheerleader and statistician. Thankfully, after about 2 years of constant work, Alan had cloned the Epo receptor [39]. The derived amino acid sequence indicated a 507-amino-acid protein with a single membrane- spanning segment but with no sequence resemblance to any protein in any database.
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Cell

ArticleExpression cloning of the murine erythropoietin receptor

Abstract

Anal. Biochem.

J. Biol. Chem.

Blood

Anal. Biochem.

Cell

Biochem. Biophys. Acta

J. Mol. Biol.

J. Biol. Chem.

FEBS Lett.

Cell

Biochem. Biophys. Res. Commun.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Biochem. Biophys. Acta

Cell

Molecular cloning of a CD28 cDNA by a high-efficiency COS cell expression system

Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease

Biochemistry

Sequences responsible for the altered erythropoietin responsiveness in spleen focus-forming virus strain SFFVp-infected cells are localized to a 678-base pair region at the 3′ end of the envelope gene

J. Virol.

A second human interleukin-2 binding protein that may be a component of high-affinity interleukin-2 receptors

Nature

Erythropoietin coming of age

N. Engl. J. Med.

Correction of the anemia of end-stage renal disease with recombinant human erythropoietin

N. Engl. J. Med.

Receptor-mediated endocytosis of low density lipoprotein in cultured cells

Meth. Enzymol.

A simple and very efficient method for generating cDNA libraries

Gene

Article
Expression cloning of the murine erythropoietin receptor