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

FEBS Letters

Capaldi, Roderick A¹  Hausrath, Andrew¹  Grüber, Gerhard¹  Sagermann, Martin¹ 

[1] Institute of Molecular Biology, University of Oregon, Eugene, OR 97403-1229, USA

关键词: F1-ATPase;    F1F0-ATP synthase;    Crystal;    DTE;    erythro-1;    4-dimercapto-2;    3-butanediol;    DTT;    dithiothreitol;    EACA;    6-aminohexanoic acid;    ECF1;    soluble portion of the Escherichia coli F1F0-ATPase;    EDTA;    ethylenediaminetetraacetic acid;    EGTA;    ethanedioxybis-(ethylamine) tetra-acetate;    MOPS;    3-(N-morpholino)propanesulfonic acid;    PAB;    p-aminobenzamidine;    PAGE;    polyacrylamide gel electrophoresis;    PEG;    polyethylene glycol;    PMSF;    phenylmethylsulfonylfluoride;    SDS;    sodium dodecyl sulfate;    TES;    N-tris[Hydroxymethyl]methyl-2-aminoethane-sulfonic acid;   

DOI  :  10.1016/S0014-5793(97)00528-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown