Evidence of a coupled mechanism between monoamine oxidase and peroxidase in the metabolism of tyramine by rat intestinal mitochondria.
The relationship between monoamine oxidase (EC 1.4.3.4; MAO) and peroxidase (EC 1.11.1.7; POD) in the metabolism of tyramine was investigated using the crude mitochondrial fraction of rat intestine. When tyramine was incubated with mitochondria, the formation of the peroxidase-catalysed oxidation product, 2,2'-dihydroxy-5,5'-bis(ethylamino)diphenyl (dityramine), identified by mass spectrometric analysis, was monitored spectrophotometrically. After an initial lag time, the formation rate of dityramine was linear up to 2 hr, amounting to 17 nmol x hr(-1) x mg protein(-1). A similar value was found for the oxidative deamination of tyramine catalysed by intestinal MAO. Either 10(-3) M clorgyline or 10(-3) M NaCN suppressed this reaction by completely inhibiting MAO or POD, respectively. In the former case, however, addition of H2O2 to the incubation mixture promptly started the reaction. Selective inhibition of MAO-A and MAO-B was achieved with 3 x 10(-7) M clorgyline and 3 x 10(-7) M deprenyl, respectively, and the formation rate of dityramine decreased in a corresponding manner. Preincubation with histamine or spermidine reduced the lag time without affecting the steady-state reaction rate. Higher levels of dityramine were also detected in vivo in rat intestine after oral administration of tyramine. These results indicate that the peroxidase-dependent metabolism of tyramine in the gut may be driven by H2O2 produced by MAO activities and that MAO-A is mainly responsible for this process, as well as for the oxidative deamination of tyramine.
Valoti
M
M
Istituto di Scienze Farmacologiche, Università di Siena, Italy.
Morón
J A
JA
Benocci
A
A
Sgaragli
G
G
Unzeta
M
M
D000818:Animals; D019281:Dimerization; D007422:Intestines / Q000201:enzymology*; D007700:Kinetics; D008297:Male; D008928:Mitochondria / Q000201:enzymology*; D008995:Monoamine Oxidase / Q000378:metabolism*; D010084:Oxidation-Reduction; D009195:Peroxidase / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D014439:Tyramine / Q000378:metabolism*
D009195:Peroxidase; D008995:Monoamine Oxidase; D014439:Tyramine