An improved purification of ECF1 and ECF1F0 by using a cytochrome bo- deficient strain of Escherichia coli facilitates crystallization of these complexes

Gerhard Grüber; Andrew Hausrath; Martin Sagermann; Roderick A. Capaldi

doi:10.1016/S0014-5793(97)00528-0

An improved purification of ECF₁ and ECF₁F₀ by using a cytochrome bo- deficient strain of Escherichia coli facilitates crystallization of these complexes

Gerhard Grüber, Andrew Hausrath, Martin Sagermann, Roderick A. Capaldi

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

12 Scopus citations

Abstract

A novel strategy, which employs a cytochrome bo-lacking strain (GO104) and a modified isolation procedure provides an effective approach for obtaining much purer preparations of ECF₁F₀ than described previously, as well as for isolating homogeneous and protein-chemically pure ECF₁. ECF₁ obtained in this way could be crystallized by vapor-diffusion using polyethylene glycol (PEG) as a precipitant in a form suitable for X-ray diffraction analysis. The crystals belong to the orthorhombic space group P2₁2₁2₁, with lattice parameters a = 110, b = 134, and c= 269 Å, and diffract to a resolution of at least 6.4 Å.

Original language	English (US)
Pages (from-to)	165-168
Number of pages	4
Journal	FEBS Letters
Volume	410
Issue number	2-3
DOIs	https://doi.org/10.1016/S0014-5793(97)00528-0
State	Published - Jun 30 1997
Externally published	Yes

Keywords

Crystal
F-ATPase
FF-ATP synthase

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

Access to Document

10.1016/S0014-5793(97)00528-0

Cite this

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title = "An improved purification of ECF1 and ECF1F0 by using a cytochrome bo- deficient strain of Escherichia coli facilitates crystallization of these complexes",

abstract = "A novel strategy, which employs a cytochrome bo-lacking strain (GO104) and a modified isolation procedure provides an effective approach for obtaining much purer preparations of ECF1F0 than described previously, as well as for isolating homogeneous and protein-chemically pure ECF1. ECF1 obtained in this way could be crystallized by vapor-diffusion using polyethylene glycol (PEG) as a precipitant in a form suitable for X-ray diffraction analysis. The crystals belong to the orthorhombic space group P212121, with lattice parameters a = 110, b = 134, and c= 269 {\AA}, and diffract to a resolution of at least 6.4 {\AA}.",

keywords = "Crystal, F-ATPase, FF-ATP synthase",

author = "Gerhard Gr{\"u}ber and Andrew Hausrath and Martin Sagermann and Capaldi, {Roderick A.}",

note = "Funding Information: We are grateful to Dr. G.B. Cox (Australian National University, Canberrra) and Dr. R. Gennis (University of Illinois) for the generous gift of pAN45 and the strain GO104, respectively. We also thank Dr. A.G.W. Leslie and Dr. J.E. Walker for their help on preliminary crystallographic experiments, and Dr. Brian Matthews for discussions and support. This research was supported by National Institutes of Health Grant HL24526 and the HHMI Grant GM20066. ",

year = "1997",

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doi = "10.1016/S0014-5793(97)00528-0",

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T1 - An improved purification of ECF1 and ECF1F0 by using a cytochrome bo- deficient strain of Escherichia coli facilitates crystallization of these complexes

AU - Grüber, Gerhard

AU - Hausrath, Andrew

AU - Sagermann, Martin

AU - Capaldi, Roderick A.

N1 - Funding Information: We are grateful to Dr. G.B. Cox (Australian National University, Canberrra) and Dr. R. Gennis (University of Illinois) for the generous gift of pAN45 and the strain GO104, respectively. We also thank Dr. A.G.W. Leslie and Dr. J.E. Walker for their help on preliminary crystallographic experiments, and Dr. Brian Matthews for discussions and support. This research was supported by National Institutes of Health Grant HL24526 and the HHMI Grant GM20066.

PY - 1997/6/30

Y1 - 1997/6/30

N2 - A novel strategy, which employs a cytochrome bo-lacking strain (GO104) and a modified isolation procedure provides an effective approach for obtaining much purer preparations of ECF1F0 than described previously, as well as for isolating homogeneous and protein-chemically pure ECF1. ECF1 obtained in this way could be crystallized by vapor-diffusion using polyethylene glycol (PEG) as a precipitant in a form suitable for X-ray diffraction analysis. The crystals belong to the orthorhombic space group P212121, with lattice parameters a = 110, b = 134, and c= 269 Å, and diffract to a resolution of at least 6.4 Å.

AB - A novel strategy, which employs a cytochrome bo-lacking strain (GO104) and a modified isolation procedure provides an effective approach for obtaining much purer preparations of ECF1F0 than described previously, as well as for isolating homogeneous and protein-chemically pure ECF1. ECF1 obtained in this way could be crystallized by vapor-diffusion using polyethylene glycol (PEG) as a precipitant in a form suitable for X-ray diffraction analysis. The crystals belong to the orthorhombic space group P212121, with lattice parameters a = 110, b = 134, and c= 269 Å, and diffract to a resolution of at least 6.4 Å.

KW - Crystal

KW - F-ATPase

KW - FF-ATP synthase

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