F0 complex of the Escherichia coli ATP synthase. Not all monomers of the subunit c oligomer are involved in F1 interaction.
The antigenic determinants of mAbs against subunit c of the Escherichia coli ATP synthase were mapped by ELISA using overlapping synthetic heptapeptides. All epitopes recognized are located in the hydrophilic loop region and are as follows: 31-LGGKFLE-37, 35-FLEGAAR-41, 36-LEGAAR-41 and 36-LEGAARQ-42. Binding studies with membrane vesicles of different orientation revealed that all mAbs bind to everted membrane vesicles independent of the presence or absence of the F1 part. Although the hydrophilic region of subunit c and particularly the highly conserved residues A40, R41, Q42 and P43 are known to interact with subunits gamma and epsilon of the F1 part, the mAb molecules have no effect on the function of F0. Furthermore, it could be demonstrated that the F1 part and the mAb molecule(s) are bound simultaneously to the F0 complex suggesting that not all c subunits are involved in F1 interaction. From the results obtained, it can be concluded that this interaction is fixed, which means that subunits gamma and epsilon do not switch between the c subunits during catalysis and furthermore, a complete rotation of the subunit c oligomer modified with mAb(s) along the stator of the F1F0 complex, proposed to be composed of at least subunits b and delta, seems to be unlikely.
Birkenhäger
R
R
Universität Osnabrück, Fachbereich Biologie/Chemie, Abteilung Mikrobiologie, Osnabrück, Germany.
Greie
J C
JC
Altendorf
K
K
Deckers-Hebestreit
G
G
D025181:ATP Synthetase Complexes; D000595:Amino Acid Sequence; D000609:Aminoacridines; D000911:Antibodies, Monoclonal / Q000276:immunology; D001665:Binding Sites; D018604:Epitope Mapping; D004926:Escherichia coli / Q000201:enzymology*; D005456:Fluorescent Dyes; D009097:Multienzyme Complexes / Q000276:immunology*; D010446:Peptide Fragments / Q000276:immunology; D017856:Phosphotransferases (Phosphate Group Acceptor) / Q000276:immunology*; D011485:Protein Binding; D006180:Proton-Translocating ATPases / Q000737:chemistry / Q000276:immunology*
D000609:Aminoacridines; D000911:Antibodies, Monoclonal; D005456:Fluorescent Dyes; D009097:Multienzyme Complexes; D010446:Peptide Fragments; C010847:9-amino-6-chloro-2-methoxyacridine; D025181:ATP Synthetase Complexes; D017856:Phosphotransferases (Phosphate Group Acceptor); D006180:Proton-Translocating ATPases