Mechanistic studies of para-substituted N,N'-dibenzyl-1,4-diaminobutanes as substrates for a mammalian polyamine oxidase.

The kinetics of oxidation of a series of para-substituted N,N'-dibenzyl-1,4-diaminobutanes by the flavoprotein polyamine oxidase from mouse have been determined to gain insight into the mechanism of amine oxidation by this member of the monoamine oxidase structural family. The k(cat)/K(m) values are maximal at pH 9, consistent with the singly charged substrate being the active form. The rate constant for flavin reduction, k(red), by N,N'-dibenzyl-1,4-diaminobutane decreases about 5-fold below a pK(a) of approximately 8; this is attributed to the need for a neutral nitrogen at the site of oxidation. The k(red) and k(cat) values are comparable for each of the N,N'-dibenzyl-1,4-diaminobutanes, consistent with rate-limiting reduction. The deuterium kinetic isotope effects on k(red) and k(cat) are identical for each of the N,N'-dibenzyl-1,4-diaminobutanes, consistent with rate-limiting cleavage of the substrate CH bond. The k(red) values for seven different para-substituted N,N'-dibenzyl-1,4-diaminobutanes correlate with a combination of the van der Waals volume and sigma value of the substrates, with rho values of -0.59 at pH 8.6 and -0.09 at pH 6.6. These results are consistent with direct transfer of a hydride from the neutral CN bond of the substrate to the flavin as the mechanism of polyamine oxidase.

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