The coupling region of F0F1 ATP synthase: binding of the hydrophilic loop of F0 subunit c to F1

• Published in: FEBS letters Vol. 431; no. 3; pp. 419 - 422
• Main Authors: Licher, T, Kellner, E, Lill, H
• Format: Journal Article
• Language: English
• Published: England 24.07.1998
• Subjects: Amino Acid Sequence; ATP synthase; ATP-Binding Cassette Transporters; ATPase; Carrier Proteins - metabolism; CF1(−δϵ), CF1 lacking subunits δ and ϵ; CF1, spinach chloroplast F1 ATPase; Coumarin, 7-diethylamino-3-(4′-maleimidylphenyl)-4-methylcoumarin; Coupling; Escherichia coli - genetics; Escherichia coli Proteins; F0F1; Fluorescein, fluorescein-5-maleimide; FRET, fluorescence resonance energy transfer; Maltose-Binding Proteins; Molecular Sequence Data; Monosaccharide Transport Proteins; PCR, polymerase chain reaction; Periplasmic Binding Proteins; Protein Binding; Proteolipid; Proton-Translocating ATPases - metabolism; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; Spectrometry, Fluorescence; Tris, N-tris-(hydroxymethyl)-aminomethane
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/S0014-5793(98)00807-2

The hydrophilic loop region of the CF0 c subunit has been expressed as a fusion with MalE in Escherichia coli. A cysteine was introduced at the C‐terminus to allow fluorophore labeling of the fusion protein. After removal of the MalE moiety, the labeled peptide was used for binding studies with fluorophore‐labeled CF1. At saturation, 1 mol peptide was bound per mol CF1. Binding was abolished after removal of subunit ϵ from CF1, and partially restored by addition of recombinant ϵ.