Influence of subunit-specific antibodies on the activity of the F0 complex of the ATP synthase of Escherichia coli. II. Effects of subunit c-specific polyclonal antibodies.
After incubation of F1-stripped everted membrane vesicles with antibodies against subunit c of the ATP synthase of Escherichia coli the proton translocation through the open F0 channel was blocked. Rebinding of F1 to those vesicles is affected by the antibody concentration used. In general, the use of F(ab')2 or Fab fragments prepared from anti-c antibodies gave similar results. However, using Fab fragments a higher amount of antigenic binding sites was necessary to block the F0 complex completely, whereas extremely low amounts of Fab fragments were necessary to inhibit the binding of F1. This can be explained by an antigenic determinant of subunit c, which is only accessible to the smaller Fab fragments with a molecular mass of approximately 50,000. Incubation of F1-containing everted membranes with anti-c antibodies showed that the binding of the antibodies resulted in a displacement of F1, while simultaneously the proton translocation through F0 has been blocked. Such a displacement can only be observed after incubation with IgG molecules or F(ab')2 fragments. Fab fragments were not able to displace the F1 part, indicating that the ability of antibodies and F(ab')2 fragments to produce cross-links is responsible for the loss of F1 from the membranes.
Deckers-Hebestreit
G
G
Universität Osnabrück, Fachbereich Biologie/Chemie, Federal Republic of Germany.
Altendorf
K
K
D000255:Adenosine Triphosphate / Q000378:metabolism; D000906:Antibodies / Q000276:immunology*; D015153:Blotting, Western; D000939:Epitopes / Q000276:immunology; D004926:Escherichia coli / Q000201:enzymology*; D006868:Hydrolysis; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*
D000906:Antibodies; D000939:Epitopes; D007140:Immunoglobulin Fab Fragments; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases