Abstract
1. 1. The fluorescence increase occurring when DPNH is bound to liver ADH has been used for determining the dissociation constant of ADH. DPNH at pH values from 5.3 to 10. The effects of formate and chloride were studied. 2. 2. The dissociation constant of ADH.DPN was determined under corresponding conditions, using as indicator the competition of DPNH and DPN for the same binding site(s). D ox, unlike D red, ascribes a univalent dissociation curve with p K′ = 7.5. The implications of this are discussed. 3. 3. Whereas good agreement was found between the present values for D red and those calculated from previous kinetic measurements, discrepancies were observed in the case of D ox. Since formate competes with ethanol for a binding site at ADH.DPN, these discrepancies can be ascribed to the formation of a ternary ADH.DPN.ethanol complex, interfering with the kinetics particularly in acid solution.
Published Version
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