Reconstitution of CF0F1 into liposomes using a new reconstitution procedure
Open Access
- 1 November 1990
- journal article
- Published by Wiley in European Journal of Biochemistry
- Vol. 193 (3), 921-925
- https://doi.org/10.1111/j.1432-1033.1990.tb19418.x
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 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.Keywords
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