The mechanism of the vanadate (V(V]-dependent oxidation of NADH was different in phosphate buffers and in phosphate-free media. In phosphate-free media (aqueous medium or HEPES buffer) the vanadyl (V(IV] generated by the direct V(V)-dependent oxidation of NADH formed a complex with V(V). In phosphate buffers V(IV) autoxidized instead of forming a complex with V(V). The generated superoxide radical (O2-) initiated, in turn, a high-rate free radical chain oxidation of NADH. Phosphate did not stimulate the V(V)-dependent NADH oxidation catalyzed by O2--generating systems. Monovanadate proved to be a stronger catalyzer of NADH oxidation as compared to polyvanadate.
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