Refolding of the isolated extracellular domain of the erythropoietin receptor.
Refolding of the soluble recombinant binding-active extracellular domain of the murine erythropoietin receptor (sEPO-R) was achieved with greater than 90% recovery either from urea/NaCI (1.5/0.5 M) or guanidine HCI (1 M). An expression plasmid encoding the extracellular plasma region of the human EPO-R (sEPO-R) was transfected into COS cells, and the sEPO-R so produced was labelled with [35S]methionine and purified by EPO-affinity chromatography on a EAH-Sepharose 4B-EPO column. Rapid rates of refolding were required for recovery of the native proteins. Refolding was evaluated by rebinding of the sEPO-R to the EPO affinity column.
Bessalle
R
R
Graduate Department of Biochemistry, Brandeis University, Waltham, MA 02154.
D'Andrea
A
A
Fasman
G
G
D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004921:Erythropoietin; D019791:Guanidine; D006146:Guanidines; D051379:Mice; D008970:Molecular Weight; D011489:Protein Denaturation; D017510:Protein Folding; D017467:Receptors, Erythropoietin / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D014162:Transfection; D014508:Urea
D006146:Guanidines; D017467:Receptors, Erythropoietin; D011994:Recombinant Proteins; D004921:Erythropoietin; D014508:Urea; D019791:Guanidine