One-step purification of Escherichia coli H+-ATPase (F0F1) and its reconstitution into liposomes with neurotransmitter transporters

Yoshinori Moriyama, Atsuko Iwamoto, Hironori Hanada, Masatomo Maeda, Masamitsu Futai

Research output: Contribution to journalArticle

114 Citations (Scopus)

Abstract

About 30% of the protein in the inner membrane of Escherichia coli strain DK8/pBWU13 is H+-ATPase (F0F1), and practically homogeneous F0F1 could be obtained by gradient centrifugation after solubilization of these membranes. The recombinant plasmid pBWU13 carries the unc operon for F0F1. When reconstituted into liposomes, F0F1 formed an ATP-dependent proton gradient and membrane potential. Proteoliposomes reconstituted with F0F1 and solubilized transporters from chromaffin granules or synaptic vesicle membranes could transport serotonin, dopamine, and norepinephrine dependent on ATP hydrolysis. F0F1 can be obtained rapidly from DK8/pBWU13, and its reconstitution into liposomes with transporters may be useful for monitoring these transporters during their purification.

Original languageEnglish
Pages (from-to)22141-22146
Number of pages6
JournalJournal of Biological Chemistry
Volume266
Issue number33
Publication statusPublished - 1991
Externally publishedYes

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Neurotransmitter Transport Proteins
Proton-Translocating ATPases
Liposomes
Escherichia coli
Purification
Adenosine Triphosphate
Chromaffin Granules
Membranes
Synaptic Membranes
Synaptic Vesicles
Operon
Centrifugation
Membrane Potentials
Protons
Dopamine
Serotonin
Norepinephrine
Hydrolysis
Plasmids
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

One-step purification of Escherichia coli H+-ATPase (F0F1) and its reconstitution into liposomes with neurotransmitter transporters. / Moriyama, Yoshinori; Iwamoto, Atsuko; Hanada, Hironori; Maeda, Masatomo; Futai, Masamitsu.

In: Journal of Biological Chemistry, Vol. 266, No. 33, 1991, p. 22141-22146.

Research output: Contribution to journalArticle

Moriyama, Yoshinori ; Iwamoto, Atsuko ; Hanada, Hironori ; Maeda, Masatomo ; Futai, Masamitsu. / One-step purification of Escherichia coli H+-ATPase (F0F1) and its reconstitution into liposomes with neurotransmitter transporters. In: Journal of Biological Chemistry. 1991 ; Vol. 266, No. 33. pp. 22141-22146.
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AU - Futai, Masamitsu

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