Functional production of the Na+ F1FO ATP synthase from Acetobacterium woodii in Escherichia coli requires the native AtpI

Autor(en): Brandt, Karsten
Mueller, Daniel B.
Hoffmann, Jan
Huebert, Christine
Brutschy, Bernd
Deckers-Hebestreit, Gabriele 
Mueller, Volker
Stichwörter: ATP synthase; AtpI; Bioenergetics; Biophysics; Cell Biology; ENERGY-CONSERVATION; Hybrid rotor; IDENTIFICATION; INTERMEDIATE STEP; Membrane enzymes; OLIGOMER; OPERON; PROTEIN; PURIFICATION; RING; Sodium transport; SUBUNIT; TRANSLOCATING F1F0-ATPASE
Erscheinungsdatum: 2013
Herausgeber: SPRINGER/PLENUM PUBLISHERS
Journal: JOURNAL OF BIOENERGETICS AND BIOMEMBRANES
Volumen: 45
Ausgabe: 1-2
Startseite: 15
Seitenende: 23
Zusammenfassung: 
The Na+ F1FO ATP synthase of the anaerobic, acetogenic bacterium Acetobacterium woodii has a unique FOVO hybrid rotor that contains nine copies of a F-O-like c subunit and one copy of a V-O-like c (1) subunit with one ion binding site in four transmembrane helices whose cellular function is obscure. Since a genetic system to address the role of different c subunits is not available for this bacterium, we aimed at a heterologous expression system. Therefore, we cloned and expressed its Na+ F1FO ATP synthase operon in Escherichia coli. A Delta atp mutant of E. coli produced a functional, membrane-bound Na+ F1FO ATP synthase that was purified in a single step after inserting a His(6)-tag to its beta subunit. The purified enzyme was competent in Na+ transport and contained the FOVO hybrid rotor in the same stoichiometry as in A. woodii. Deletion of the atpI gene from the A. woodii operon resulted in a loss of the c ring and a mis-assembled Na+ F1FO ATP synthase. AtpI from E. coli could not substitute AtpI from A. woodii. These data demonstrate for the first time a functional production of a FOVO hybrid rotor in E. coli and revealed that the native AtpI is required for assembly of the hybrid rotor.
ISSN: 0145479X
DOI: 10.1007/s10863-012-9474-8

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