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.