Активизация пируватдегидрогеназного комплекса и ингибирование цикла Кребса и дыхания митохондрий избытком пирувата

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Аннотация

В экспериментах на изолированных митохондриях печени крыс показано, что пируват (10–30 мМ) в присутствии L-глутамата вызывает концентрационно-зависимое ингибирование дыхания, активированного ADP. Дыхание реактивируется 3 мМ L-малата. Оба эффекта воспроизводятся в присутствии D,L-ацетилкарнитина (АсCar), что указывает на важную роль ацетилКоА (AcCoA) в регуляции реакций цикла Кребса. При окислении пирувата скорость дыхания снижается в течение нескольких сотен секунд. Эффект воспроизводится в присутствии дихлорацетата (DCA), ингибитора киназы пируватдегидрогеназы (PDK) и не наблюдается при избытке АсCar, что указывает на дефосфорилирование пируватдегидрогеназы (PDH) при ингибировании PDK пируватом (+ADP) или DCA. Эффекты пирувата и АсСar зависят от времени преинкубации митохондрий в состоянии 2. Эксперименты на замороженных/размороженных митохондриях показывают, что преинкубация митохондрий с пируватом восстанавливает активность PDH и подавляет активность α-кетоглутаратдегидрогеназы (α-KGDH), регистрируемых по флуоресценции NADH. Таким образом, в качестве возможного механизма ингибирования дыхания пируватом можно предположить комбинированный механизм, сочетающий 1) аллостерическое ингибирование цитратсинтазы избытком AcCoA при низких концентрациях оксалоацетата и α-KGDH с возможным участием ацетоацетилКоА; 2) медленное ацетилирование α-KGDH и других ферментов цикла избытком AcCoA при медленной реактивации PDH пируватом.

Об авторах

В. В. Дынник

Институт теоретической и экспериментальной биофизики РАН

Email: dynnik@rambler.ru
Пущино, 142290 Россия

Е. В. Гришина

Институт теоретической и экспериментальной биофизики РАН

Пущино, 142290 Россия

Н. И. Федотчева

Институт теоретической и экспериментальной биофизики РАН

Пущино, 142290 Россия

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