Characterization of monoclonal antibodies against mitochondrial F1-ATPase.

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Abstract
Four stable murine hybridoma clones producing homogeneous antibodies against pig heart mitochondrial F1-ATPase were established. All antibodies exhibit specific binding to F1-ATPase. Three of them interact with the .beta. subunit and one binds strongly to the .alpha. and barely to the .beta. subunit. All of them exhibit linear Scatchard plots and high binding affinities, with Kd between 4.7 .times. 10-8 M and 6.5 .times. 10-10 M. The minimal number of moles of IgG bound at saturation per mole of immobilized F1-ATPase is 2.2 for the anti-.alpha.-subunit antibody and 2.5 for one anti-.beta.-subunit antibody. This suggests that more than 2 copies of the .alpha. and .beta. subunits are present in the enzyme. Two antibodies seem to recognize F1-ATPase equally before or after denaturation with sodium dodecyl sulfate. The 2 other antibodies exhibited a much higher affinity for the nondissociated enzyme, indicating that they are very sensitive to a specific conformation of the enzyme.