ABSTRACT
It has been shown that monoclonal antibody (mAb) F7F10 raised against pyruvate dehydrogenase component (E1) of pigeon breast muscle pyruvate dehydrogenase complex (PDC) has no influence on the E1 activity, measured in the system with artificial oxidants. However it inhibited the full NAD+ and coenzyme A dependent activity of PDC. The competition of the F7F10 antibody with the E2 component of PDC for the binding with E1 was revealed by immunoenzymatic and kinetic analysis. It is suggested that F7F10 mAb interacts with an antigenic determinant, located in the immediate vicinity of or overlapping with the E1 region, responsible for the interaction with the E2 component of PDC.
Subject(s)
Antibodies, Monoclonal/pharmacology , Epitopes/immunology , Pyruvate Dehydrogenase Complex/immunology , Animals , Antibodies, Monoclonal/metabolism , Binding Sites, Antibody , Binding, Competitive , Columbidae , Epitopes/metabolism , Humans , Ketoglutarate Dehydrogenase Complex/drug effects , Ketoglutarate Dehydrogenase Complex/immunology , Kinetics , Muscles/enzymology , Myocardium/enzymology , NAD/metabolism , Pyruvate Dehydrogenase Complex/drug effects , Pyruvate Dehydrogenase Complex/metabolism , Thiamine Pyrophosphate/immunology , Thiamine Pyrophosphate/metabolismABSTRACT
The monoclonal antibody F7F10 against the E1 component of the pigeon breast muscle pyruvate dehydrogenase complex has been produced. The dissociation constant of the E1-mAb F7F10 complex was determined to be 5.93 x 10(-8)M. The cross-reaction of the mAb with the E1 components of the pyruvate dehydrogenase complexes from various species (including human) was established. The competitive solid-phase immunoenzyme assay of the E1 component and PDC concentrations has been developed.
