Immunochemical analysis of Micrococcus lysodeikticus (luteus) F1-ATPase and its subunits.
The F1-ATPase from Micrococcus lysodeikticus has been purified to 95% protein homogeneity in this laboratory and as all other bacterial F1S, possesses five distinct subunits with molecular weights ranging from 60 000 to 10 000 (Huberman, M. and Salton, M.R.J. (1979) Biochim. Biophys. Acta 547, 230-240). In this communication, we demonstrate the immunochemical reactivities of antibodies to native and SDS-dissociated subunits with the native and dissociated F1-ATPase and show that: (1) the antibodies generated to the native or SDS-dissociated subunits react with the native molecule; (2) all of the subunits comprising the F1 are antigenically unique as determined by crossed immunoelectrophoresis and the Ouchterlony double-diffusion techniques; (3) antibodies to the SDS-denatured individual delta- and epsilon-subunits can be used to destabilize the interaction of these specific subunits with the rest of the native F1; and (4) all subunit antibodies as well as anti-native F1 were found to inhibit ATPase activity to varying degrees, the strongest inhibition being seen with antibodies to the total F1 and anti-alpha- and anti-beta-subunit antibodies. The interaction of specific subunit antibodies may provide a new and novel way to study further and characterize the catalytic portions of F1-ATPases and in general may offer an additional method for the examination of multimeric proteins.
D000936:Antigen-Antibody Complex; D007106:Immune Sera; D007122:Immunoelectrophoresis; D046911:Macromolecular Substances; D008837:Micrococcus / Q000201:enzymology*; D008970:Molecular Weight; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors / Q000276:immunology / Q000302:isolation & purification*
D000936:Antigen-Antibody Complex; D007106:Immune Sera; D046911:Macromolecular Substances; D006180:Proton-Translocating ATPases