Reconstitution of CF0F1 into liposomes using a new reconstitution procedure

Peter Richard, Jean Louis Rigaud, Peter Gräber

Research output: Contribution to journalArticleScientificpeer-review

71 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)921-925
Number of pages5
JournalEuropean Journal of Biochemistry
Volume193
Issue number3
DOIs
Publication statusPublished - Jan 1990
MoE publication typeNot Eligible

Fingerprint

Liposomes
Adenosine Triphosphate
Chloroplast Proton-Translocating ATPases
Proton-Translocating ATPases
Octoxynol
Phosphatidylcholines
Detergents
Permeability
Evaporation
Proteins
Membranes

Cite this

Richard, Peter ; Rigaud, Jean Louis ; Gräber, Peter. / Reconstitution of CF0F1 into liposomes using a new reconstitution procedure. In: European Journal of Biochemistry. 1990 ; Vol. 193, No. 3. pp. 921-925.
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Reconstitution of CF0F1 into liposomes using a new reconstitution procedure. / Richard, Peter; Rigaud, Jean Louis; Gräber, Peter.

In: European Journal of Biochemistry, Vol. 193, No. 3, 01.1990, p. 921-925.

Research output: Contribution to journalArticleScientificpeer-review

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