The transmembrane domain of subunit b of the Escherichia coli F1F0 ATP synthase is sufficient for H+-translocating activity together with subunits a and c

Autor(en): Greie, JC
Heitkamp, T
Altendorf, K 
Stichwörter: 2ND STALK; BINDING; Biochemistry & Molecular Biology; CROSS-LINKING; DELTA-SUBUNIT; EPSILON-SUBUNIT; Escherichia coli; F-0 COMPLEX; F1FO ATP synthase; ORGANIZATION; PROTEINS; proton translocation; RECONSTITUTION; subunit b; TRANSPORT
Erscheinungsdatum: 2004
Volumen: 271
Ausgabe: 14
Startseite: 3036
Seitenende: 3042
Subunit b is indispensable for the formation of a functional H+-translocating F-O complex both in vivo and in vitro. Whereas the very C-terminus of subunit b interacts with F-1 and plays a crucial role in enzyme assembly, the C-terminal region is also considered to be necessary for proper reconstitution of F-O into liposomes. Here, we show that a synthetic peptide, residues 1-34 of subunit b (b(1-34)) [Dmitriev, O., Jones, P.C., Jiang, W. & Fillingame, R.H. (1999) J. Biol. Chem.274, 15598-15604], corresponding to the membrane domain of subunit b was sufficient in forming an active F-O complex when coreconstituted with purified ac subcomplex. H+ translocation was shown to be sensitive to the specific inhibitor N,N'-dicyclohexylcarbodiimide, and the resulting F-O complexes were deficient in binding of isolated F-1. This demonstrates that only the membrane part of subunit b is sufficient, as well as necessary, for H+ translocation across the membrane, whereas the binding of F-1 to F-O is mainly triggered by C-terminal residues beyond Glu34 in subunit b. Comparison of the data with former reconstitution experiments additionally indicated that parts of the hydrophilic portion of the subunit b dimer are not involved in the process of ion translocation itself, but might organize subunits a and c in F-O assembly. Furthermore, the data obtained functionally support the monomeric NMR structure of the synthetic b(1-34).
ISSN: 00142956
DOI: 10.1111/j.1432-1033.2004.04235.x

Show full item record

Google ScholarTM