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 ΔpH/ΔΨ. 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.
|Number of pages||5|
|Journal||European Journal of Biochemistry|
|Publication status||Published - Jan 1990|
|MoE publication type||Not Eligible|