Purification of the proton pumping ATPase from plant plasma membranes.
The plasma membrane ATPase from oat roots has been purified near homogeneity by a simple procedure. Plasma membranes isolated from sucrose gradients are first extracted with Triton X-100 and KC1 and the residue solubilized with lysolecithin. Rate-zonal centrifugation in a vertical rotor with a glycerol gradient results in a preparation of very high specific activity (6 mumoles min-1 mg protein-1 at 30 degrees C) and where over 70% of the protein corresponds to a polypeptide of about 100 kilodaltons previously identified as the ATPase. The purified enzyme could be reconstituted in proteoliposomes catalyzing ATP-driven proton transport sensitive to vanadate.
D002384:Catalysis; D002462:Cell Membrane / Q000201:enzymology; D002499:Centrifugation, Density Gradient; D002523:Edible Grain / Q000201:enzymology*; D006180:Proton-Translocating ATPases / Q000302:isolation & purification* / Q000378:metabolism
D006180:Proton-Translocating ATPases