Evidence from immunological studies of structure-mechanism relationship of F1 and F1F0

Abstract

Monoclonal and polyclonal antibodies directed against peptides of F1-ATPase of 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 beta subunit of epitopes in the proximity of, or in the catalytic site, gives information on this site. The use of anti-alpha monoclonal antibodies has shown asymmetry of alpha in the complex as already shown for beta. In addition, the involvement of alpha 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.

Intrinsic tryptophan fluorescence of Schizosaccharomyces pombe mitochondrial F1-ATPase. A powerful probe for phosphate and nucleotide interactions

“…Nucleotideand phosphate-induced conformational changes of the enzyme have been characterized in our laboratory by a number of approaches including accessibility to proteases (Di Pietro et al" 1983), circular dichroism (Roux et al, 1984), and reactivity to various chemical reagents (Di Pietro et al, 1979;Fellous et al, 1984;Faison et al, 1986) and monoclonal antibodies (Gautheron & Godinot, 1988).…”