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Characterization of the Reaction Intermediates in a Model Oxidative Phosphorylation Reaction

From the ¹ From the Kline Chemistry Laboratory, Yale University, New Haven, Connecticut 06520, and the Department of Chemistry, Temple University, Philadelphia, Pennsylvania 19122

Methods are described to separate or trap and subsequently characterize the intermediates in the previously reported model oxidative phosphorylation reaction in which ATP is produced from orthophosphate and AMP or ADP during the oxidation of a ferrohemochrome by molecular oxygen in N, N-dimethylacetamide solution containing an excess of imidazole.

By using ¹⁴C-labeled imidazole and ³²P-labeled P_i, it is shown that the "energy-rich" intermediate is indistinguishable from 1-phosphoimidazole in its chromatographic, electrophoretic, and chemical properties. Similar phosphorylated intermediates can be formed with 2-methylimidazole and 2,4-dimethylimidazole, but not with 1,2-dimethylimidazole.

A molecular mechanism of this model oxidative phosphorylation reaction is suggested.

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