Exploration of δ‐subunit interactions in beef heart mitochondrial F1 ‐ATPase by monoclonal antibodies
- 13 March 1989
- journal article
- research article
- Published by Wiley in FEBS Letters
- Vol. 245 (1-2) , 223-228
- https://doi.org/10.1016/0014-5793(89)80226-1
Abstract
Three monoclonal antibodies (mAbs) recognizing distinct epitopes on the δ‐subunit of beef heart mitochondrial F1‐ATPase were studied for their reactivity towards the δ‐subunit both in isolated F1 and in the F0‐F1 complex of submitochondrial particles. Two of the antibodies termed mAb δ 195 and mAb δ239 had free access to δ in F1 and the F0‐F1 complex. Partial hindrance was observed for the third antibody mAb δ22. By a double antibinding assay, it was found that the binding sites for mAb δ195 and mAb δ239 were close to each other and possibly overlapping. Mapping studies conducted with the isolated δ‐subunit showed that mAb δ195 and mAb δ239 interacted with the N‐terminal portion of δ extending from Ala‐1 to Met‐16, whereas mAb δ22 interacted with the fragment spanning Ser‐17‐Glu‐68. It was concluded that the Ala‐1‐Met‐16 segment of the δ‐subunit in F1 and the F0‐F1 complex is freely accessible from the outside, whereas the Ser‐17‐Glu‐68 segment of δ is partially hidden, possibly as a result of interactions with other subunits.Keywords
This publication has 26 references indexed in Scilit:
- Isolation and characterisation of monoclonal antibodies against hydrophobic membrane subunit 9 of the yeast mitochondrial H+-ATPaseBiochimica et Biophysica Acta (BBA) - Bioenergetics, 1988
- Primary structure and subunit stoichiometry of F1-ATPase from bovine mitochondriaJournal of Molecular Biology, 1985
- Monoclonal antibodies to mitochondrial coupling factor BFEBS Letters, 1985
- Monoclonal antibodies to mitochondrial F1-ATPase and oligomycin sensitivity conferring protein (OSCP). Tools for recognition of well conserved and essential antigenic sitesBiochemical and Biophysical Research Communications, 1984
- Monoclonal antibodies against subunits of yeast mitochondrial H+-ATPaseBiochimica et Biophysica Acta (BBA) - Bioenergetics, 1984
- Use of monoclonal antibodies in immuno-electron microscopy for the determination of subunit stoichiometry in oligomeric enzymes: There are three α-subunits in the F1-ATPase of Escherichia coliJournal of Molecular Biology, 1984
- Aurovertin binding sites on beef heart mitochondrial F1-ATPase. Study with [14C]aurovertin D of the binding stoichiometry and of the interaction between aurovertin and the natural ATPase inhibitor for binding to F1Biochemistry, 1983
- Isolation of pure IgG1, IgG2a and IgG2b immunoglobulins from mouse serum using protein A-SepharoseImmunochemistry, 1978
- A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye bindingAnalytical Biochemistry, 1976
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970