Evidence from immunological studies of structure-mechanism relationship of F1 and F1F0 |
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Authors: | Danièle C Gautheron Catherine Godinot |
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Institution: | (1) Laboratoire de Biologie et Technologie des Membranes du CNRS, Université Claude Bernard de Lyon, 69622 Villeurbanne Cedex, France |
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Abstract: | Monoclonal and polyclonal antibodies directed against peptides of F1-ATPase or F1F0-ATPase synthase provide new and efficient tools to study structure-function relationships and mechanisms of such complex membrane enzymes. This review summarizes the main results obtained using this approach. Antibodies have permitted the determination of the nature of subunits involved in the complex, their stoichiometry, their organization, neighboring interactions, and vectorial distribution within or on either face of the membrane. Moreover, in a few cases, amino acid sequences exposed on a face of the membrane or buried inside the complex have been identified. Antibodies are very useful for detecting the role of each subunit, especially for those subunits which appear to have no direct involvement in the catalytic mechanism. Concerning the mechanisms, the availability of monoclonal antibodies which inhibit (or activate) ATP hydrolysis or ATP synthesis, which modify nucleotide binding or regulation of activities, which detect specific conformations, etc. brings many new ways of understanding the precise functions. The specific recognition by monoclonal antibodies on the subunit of epitopes in the proximity of, or in the catalytic site, gives information on this site. The use of anti- monoclonal antibodies has shown asymmetry of in the complex as already shown for . In addition, the involvement of with respect to nucleotide site cooperativity has been detected. Finally, the formation of F1F0-antibody complexes of various masses, seems to exclude the functional rotation of F1 around F0 during catalysis.Abbreviations IF1
natural protein inhibitor of the ATPase-ATP synthase
- OSCP
oligomycin sensitivity-conferring protein
- DCCD
dicyclohexylcarbodiimide
- SDS-PAGE
sodium dodecylsulfate polyacrylamide gel electrophoreses
- F1
F1-ATPase, coupling factor F1 of ATPase
- F1F0
F1F0-ATP synthase, ATPase-ATP synthase complex |
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Keywords: | Monoclonal polyclonal antibodies F1-ATPase F1-F0-ATP synthase immunological approach structure assembly mechanisms regulation |
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