title: Reconstitution of CF0F1 into liposomes using a new reconstitution procedure.
abstract: The H(+)-ATPase (ATP synthase) from chloroplasts was isolated, purified and reconstituted into phosphatidylcholine/phosphatidic-acid liposomes. Liposomes prepared by reverse-phase evaporation were treated with various amounts of Triton X-100 and protein incorporation was studied at each step of the solubilization process. After detergent removal by SM2-Biobeads, the activities of the resulting proteoliposomes were measured indicating that the most efficient reconstitution was obtained by insertion of the protein into preformed, detergent-saturated liposomes. The conditions for the reconstitution were optimized with regard to ATP synthesis driven by an artificially generated delta pH/delta psi. An important benefit of the new reconstituted CF0F1 liposomes is the finding that the rate of ATP synthesis remains constant up to 10 s, indicating a low basal membrane permeability.
authors: Max-Volmer-Institut, Technische Universität Berlin, Federal Republic of Germany.
authors: P
authors: P
authors: Richard
authors: J L
authors: JL
authors: Rigaud
authors: P
authors: P
authors: Gräber
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D002736:Chloroplasts / Q000201:enzymology*; D003902:Detergents; D007700:Kinetics; D008081:Liposomes; D017830:Octoxynol; D010712:Phosphatidic Acids; D010713:Phosphatidylcholines; D011092:Polyethylene Glycols; D006180:Proton-Translocating ATPases / Q000378:metabolism*
chemical_list: D003902:Detergents; D008081:Liposomes; D010712:Phosphatidic Acids; D010713:Phosphatidylcholines; D011092:Polyethylene Glycols; D000255:Adenosine Triphosphate; D017830:Octoxynol; D006180:Proton-Translocating ATPases