Молекулярный механизм генерации мембранного потенциала цитохром с оксидазой
Диссертация
В концевом участке дыхательных цепей аэробных организмов образование протон-движущей силы обеспечивается в результате работы терминальных оксидаз, катализирующих восстановление кислорода цитохромом с или хинолом. Основная подгруппа этих ферментов, включающая цитохромоксидазу (ЦО) из митохондрий, образует суперсемейство структурно-родственных гем-медных оксидаз. Отличительной особенностью… Читать ещё >
Содержание
- СПИСОК СОКРАЩЕНИЙ
- 1. ОБЗОР ЛИТЕРАТУРЫ
- 1. 1. Общая характеристика цитохромоксидазы
- 1. 2. Кристаллическая структура цитохромоксидазы. 1О
- 1. 3. Редокс-центры и субъединицы каталитического ядра цитохромоксидазы
- 1. 4. Пути проведения протонов, переноса молекулярного кислорода и воды
- 1. 5. Физико-химические свойства цитохромоксидазы
- 1. 6. Подходы к изучению переноса зарядов в цитохромоксидазе с временным разрешением
- 1. 7. Промежуточные состояния в каталитическом цикле цитохромоксидазы
- 1. 8. Представления о сопряженном протонном транспорте в цитохромоксидазе, базирующееся на стационарных измерениях
- 1. 9. Структурно-функциональные особенности цитохромоксидазы Ъаз из ТлЬегторкИт
- 2. МАТЕРИАЛЫ И МЕТОДЫ ИССЛЕДОВАНИЯ
- 2. 1. Митохондриальная цитохромоксидаза
- 2. 2. Штамм КкойоЬа^ег яр}1аего1с1е
- 2. 3. Выращивание клеток и получение мембран из клеток Я. яркаегогсЗея
- 2. 4. Изоляция цитохромоксидазы ааз из Кхркаег oid. es
- 2. 5. Определение активности цитохромоксидазы ааз из К.5ркаего1йе
- 2. 6. Цитохромоксидаза Ъаъ из ТЪегтш 1кегторЫ1ш
- 2. 7. Встраивание цитохромоксидазы в протеолипосомы
- 2. 8. Электрометрические измерения с использованием трис (бипиридил)рутения
- 2. 9. Электрометрические измерения с ипользованием метода «флоу-флэш»
- 2. 10. Спектрофотометрические кинетические измерения на одиночной длине волны
- 2. 11. Разрешенная во времени регистрация спектров ЦО в оптическом диапазоне в сочетании с методом «флоу-флэш»
- 2. 12. Обработка данных
- 3. РЕЗУЛЬТАТЫ ИССЛЕДОВАНИЯ
- 3. 1. Генерация мембранного потенциала, сопряженная с одноэлектронными переходами в каталитическом цикле цитохромоксидазы
- 3. 1. 1. Электрогенные события, сопряженные с переносом на кислород 4-го электрона в каталитическом цикле цитохромоксидазы (Р—>0 переход)
- 3. 1. 2. Электрогенные события, сопряженные с переносом на кислород 1-го и 3-го электронов в каталитическом цикле цитохромоксидазы (фотовосстановление состояний О и Р)
- 3. 2. Кинетика переноса электрона через гемовые центры в одноэлектронных переходах каталитического цикла цитохромоксидазы
- 3. 3. Внутрибелковые пути переноса протонов в ходе одноэлектронных переходов каталитического цикла цитохромоксидазы
- 3. 3. 1. Электрогенная активность цитохромоксидазы Шз из ШюйоЪаМег яр}гаего1с1е$ дикого типа
- 3. 3. 2. Роль входных протонных Б и К каналов в каталитическом цикле цитохромоксидазы
- 3. 4. Артикуляция внутрибелкового перемещения протонов в одноэлектронном переходе Р—>0 цитохромоксидазы
- 3. 4. 1. Стехиометрия сопряженного электрогенного переноса протонов в одноэлектронном переходе Р—>
- 3. 4. 2. Последовательность проведения помпируемого и субстратного протонов в переходе Р—"0 цитохромоксидазы
- 3. 4. 3. Высвобождение протона на внешнюю сторону мембраны. Ионы цинка — как ингибитор электрогенного переноса протонов в цитохромоксидазе
- 3. 5. Особенности одноэлектронных переходов в каталитическом цикле цитохромоксидазы Ъаз из ТлЬ. егторЫ1т
- 3. 5. 1. Электрогенные события, сопряженные с переносом на кислород 1-го электрона в каталитическом цикле цитохромоксидазы Ъаз (фотовосстановление состояний О)
- 3. 5. 2. Интермедиаты каталитического цикла, разрешаемые при окислении кислородом восстановленной цитохромоксидазы Ъат, из ТЛкегторкИт в режиме одного оборота
- 3. 5. 3. Стадии генерации мембранного потенциала, сопряженные с окислением восстановленной Ъаз оксидазы из ТлкегторЫliis кислородом в режиме одного оборота
- 3. 1. Генерация мембранного потенциала, сопряженная с одноэлектронными переходами в каталитическом цикле цитохромоксидазы
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