Solubilization and reconstitution of a lysosomal H+-pump.
H+-ATPase was solubilized from the lysosomal membranes of rat liver with lysophosphatidylcholine. The detergent was removed with Biobeads SM2, and the proteoliposomes were prepared by rapid dilution of a mixture of the solubilized ATPase, octyl-beta-D-glucopyranoside and liposomes. The proteoliposomes exhibited ATP-dependent fluorescence quenching of quinacrine and bis(3-phenyl-5-oxoisoxasol-4-yl)pentamethine oxonol. These results suggested that the electrogenic lysosomal H+-pump was solubilized and reconstituted into proteoliposomes.
D000818:Animals; D007425:Intracellular Membranes / Q000201:enzymology*; D007700:Kinetics; D008081:Liposomes; D008099:Liver / Q000201:enzymology*; D008247:Lysosomes / Q000201:enzymology*; D006180:Proton-Translocating ATPases / Q000302:isolation & purification / Q000378:metabolism; D051381:Rats
D008081:Liposomes; D006180:Proton-Translocating ATPases