Stoichiometry of the topa quinone biogenesis reaction in copper amine oxidases

The stoichiometry of the topa quinone biogenesis reaction in phenylethylamine oxidase from Arthrobacter globiformis (AGAO) has been determined. We have shown that the 6e(-) oxidation of tyrosine to topa quinone (TPQ) consumes 2 mol of O-2 and produces 1 mol of H2O2/mol of TPQ formed. The rate of H2O2 production is first-order (k(obs) = 1.0 +/- 0.2 min(-1)), a rate only slightly lower than the rate of TPQ formation directly determined previously (k(obs) = 1.5 +/- 0.2 min(-1)). This gives the following net reaction stoichiometry for TPQ biogenesis: E-Tyr + 2O(2) --> E-TPQ + H2O2. This stoichiometry is in agreement with recently proposed mechanisms for T-PQ biogenesis, and rules out several possible alternatives.