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
This publication has 19 references indexed in Scilit:
- Mechanisms of membrane protein insertion into liposomes during reconstitution procedures involving the use of detergents. 2. Incorporation of the light-driven proton pump bacteriorhodopsinBiochemistry, 1988
- Mechanisms of membrane protein insertion into liposomes during reconstitution procedures involving the use of detergents. 1. Solubilization of large unilamellar liposomes (prepared by reverse-phase evaporation) by Triton X-100, octyl glucoside, and sodium cholateBiochemistry, 1988
- The rate of ATP synthesis catalyzed by reconstituted CF0F1-liposomes: Dependence on ΔpH and ΔψBiochimica et Biophysica Acta (BBA) - Bioenergetics, 1987
- ATP synthesis by ATPase proteoliposomes from the thermophilic cyanobacterium Synechococcus 6716 by ionophore-induced electric potentials and proton gradientsBiochimica et Biophysica Acta (BBA) - Bioenergetics, 1985
- The rate of ATP synthesis by reconstituted CF0F1 liposomesBiochimica et Biophysica Acta (BBA) - Bioenergetics, 1985
- Kinetics of Proton‐Transport‐Coupled ATP Synthesis in Chloroplasts. Activation of the ATPase by an Artificially Generated ΔpH and ΔψBerichte der Bunsengesellschaft für physikalische Chemie, 1984
- Use of liposomes for reconstitution of biological functionsBiochimica et Biophysica Acta (BBA) - Reviews on Biomembranes, 1982
- Phospholipase D from Savoy cabbage: purification and preliminary kinetic characterizationBiochemistry, 1979
- Assay of proteins in the presence of interfering materialsAnalytical Biochemistry, 1976
- A simple procedure for removal of triton X-100 from protein samplesAnalytical Biochemistry, 1973