Solubilization and characterization of erythropoietin receptor from transplantable mouse erythroblastic leukemic cells.
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.
Hitomi
K
K
Department of Food Science and Technology, Faculty of Agriculture, Kyoto University, Japan.
Masuda
S
S
Ito
K
K
Ueda
M
M
Sasaki
R
R
D000818:Animals; D002462:Cell Membrane / Q000032:analysis; D011232:Chemical Precipitation; D002793:Cholic Acids; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D004921:Erythropoietin / Q000378:metabolism; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism*; D051379:Mice; D008809:Mice, Inbred C3H; D009368:Neoplasm Transplantation; D011092:Polyethylene Glycols; D011956:Receptors, Cell Surface / Q000032:analysis* / Q000378:metabolism; D017467:Receptors, Erythropoietin; D012995:Solubility; D013388:Succinimides
D002793:Cholic Acids; D003432:Cross-Linking Reagents; D011956:Receptors, Cell Surface; D017467:Receptors, Erythropoietin; D013388:Succinimides; D004921:Erythropoietin; D011092:Polyethylene Glycols; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate; C019358:disuccinimidyl suberate