Stability of [3H]ouabain binding to the (Na+ + K+)-ATPase solubilized with C12E8.
The (Na+ + K+)-ATPase from dog kidney and partially purified membranes from HK dog erythrocytes were labeled with [3H]ouabain, solubilized with C12E8 and analyzed by HPLC through a TSK-GEL G3000SW column in the presence of C12E8, Mg2+, HPO4(2-) and glycerol at 20-23 degrees C. The peaks of [3H]ouabain bound to the enzyme from dog kidney and HK dog erythrocyte membranes corresponded to each other with apparent molecular weights of 470 000-490 000. In addition, these bindings of [3H]ouabain to the (Na+ + K+)-ATPase were observed to be stable at 20-23 degrees C for at least 18 h after the solubilization.
Inaba
M
M
Maede
Y
Y
D000818:Animals; D001665:Binding Sites; D002851:Chromatography, High Pressure Liquid; D003902:Detergents; D004285:Dogs; D004910:Erythrocyte Membrane / Q000201:enzymology; D007668:Kidney / Q000201:enzymology; D008861:Microsomes / Q000201:enzymology; D010042:Ouabain / Q000378:metabolism*; D011092:Polyethylene Glycols; D000254:Sodium-Potassium-Exchanging ATPase / Q000302:isolation & purification* / Q000378:metabolism; D012995:Solubility
D003902:Detergents; C017272:dodecyloctaethyleneglycol monoether; D011092:Polyethylene Glycols; D010042:Ouabain; D000254:Sodium-Potassium-Exchanging ATPase