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

Y. Moriyama; A. Iwamoto; H. Hanada; M. Maeda; M. Futai

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

Y. Moriyama, A. Iwamoto, H. Hanada, M. Maeda, M. Futai

School of Pharmaceutical Sciences

Research output: Contribution to journal › Article › peer-review

121 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 language	English
Pages (from-to)	22141-22146
Number of pages	6
Journal	Journal of Biological Chemistry
Volume	266
Issue number	33
Publication status	Published - 1991

ASJC Scopus subject areas

Biochemistry
Molecular Biology
Cell Biology

Cite this

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

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.",

author = "Y. Moriyama and A. Iwamoto and H. Hanada and M. Maeda and M. Futai",

year = "1991",

language = "English",

volume = "266",

pages = "22141--22146",

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T1 - One-step purification of Escherichia coli H+-ATPase (F0F1) and its reconstitution into liposomes with neurotransmitter transporters

AU - Moriyama, Y.

AU - Iwamoto, A.

AU - Hanada, H.

AU - Maeda, M.

AU - Futai, M.

PY - 1991

Y1 - 1991

N2 - 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.

AB - 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.

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

Abstract

ASJC Scopus subject areas

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