Subunit a of the E-coli ATP synthase: reconstitution and high resolution NMR with protein purified in a mixed polarity solvent

Autor(en): Dmitriev, OY
Altendorf, K 
Fillingame, RH
Stichwörter: ATP synthase; Biochemistry & Molecular Biology; Biophysics; Cell Biology; chloroform-methanol-water solvent; COMPLEX; CROSS-LINKING; H+; INTEGRAL MEMBRANE-PROTEIN; membrane protein purification; MOLECULAR ARCHITECTURE; MULTIDRUG TRANSPORTER; proton translocation; PURIFICATION; ROTARY MOTOR; SPECTROSCOPY; STOICHIOMETRY; subunit alpha; TROSY
Erscheinungsdatum: 2004
Herausgeber: WILEY
Journal: FEBS LETTERS
Volumen: 556
Ausgabe: 1-3
Startseite: 35
Seitenende: 38
Zusammenfassung: 
Subunit a of the Escherichia coli ATP synthase, a 30 kDa integral membrane protein, was purified to homogeneity by a novel procedure incorporating selective extraction into a monophasic mixture of chloroform, methanol and water, followed by Ni-NTA chromatography in the mixed solvent. Pure subunit a was reconstituted with subunits b and c and phospholipids to form a functional proton-translocating unit. Nuclear magnetic resonance (NMR) spectra of the pure subunit a in the mixed solvent show good chemical shift dispersion and demonstrate the potential of the solvent mixture for NMR studies of the large membrane proteins that are currently intractable in aqueous detergent solutions. (C) 2003 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
ISSN: 18733468
DOI: 10.1016/S0014-5793(03)01360-7

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