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

FEBS Letters

Dmitriev, Oleg Y.²  Altendorf, Karlheinz¹  Fillingame, Robert H.² 

[1] Fachbereich Biologie/Chemie, Universität Osnabrück, Barbarastr. 11, 49069 Osnabrück, Germany;Department of Biomolecular Chemistry, University of Wisconsin Medical School, 1300 University Avenue, Madison, WI 53706-1532, USA

关键词: ATP synthase;    Subunit a;    Membrane protein purification;    Chloroform–methanol–water solvent;    Proton translocation;    TROSY;   

DOI  :  10.1016/S0014-5793(03)01360-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown