Конформационные особенности молекулы рибулозо-1, 5-дифосфаткарбоксилазы и её активного центра, модифицированных спиновой меткой
Диссертация
Измеренное время корреляции больших субьединиц РДФ-азы (при-i близительно равное 400 нсек) и исследование взаимодействия больших и малых субьединиц с субстратом, его аналогом и активатором, позволяет сделать заключение, что большие субьединицы в отсутствии малых субьединиц агрегируют и образуют высокомолекулярные конгломераты по размерам близкие к нативному ферменту. Отсюда можно сделать вывод… Читать ещё >
Содержание
- Глава I. ОБЗОР ЛИТЕРАТУРЫ
- 1. 1. Физико-химические свойства рибулозо-1,5-дифосфат-карбоксилазы
- 1. 2. Каталитические свойства РДФ-азы
- 1. 3. Химическая природа групп активного центра РДФ-азы
- 1. 4. Особенности метода спиновых меток
- 1. 5. Зависимость параметров спектров ЭПР иминоксильных радикалов от полярности окружения
- 1. 6. Исследование белков с помощью метода спиновых меток
- ЭКСПЕРИМЕНТАЛЬНАЯ ЧАСТ
- Глава 2. МАТЕРИАЛЫ И МЕТОДЫ ИССЛЕДОВАНИЯ
- 2. 1. Выделение белка и его модификация спин-меткой
- 2. 2. Получение спин-меченых субьединиц рибулозо-I, 5-ди-фосфаткарбоксилазы
- 2. 3. Методика обработки носителей для иммобилизации рибулозо-1,5-дифосфаткарбоксилазы
- Глава 3. ИССЛЕДОВАНИЕ СПИН-МЕЧЕНОЙ РИБУЛ030−1,5-ДИФОСФАТ КАРБОКСИЛАЗЫ
- 3. 1. Идентификация места присоединения спин-меток на молекуле РДФ-азы
- 3. 2. Анализ спектра ЭПР спин-меченой РДФ-азы. Измерение времени корреляции макромолекулы РДФ-азы
- 3. 3. Изменение конформации молекулы РДФ-азы при взаимодействии с субстратами и активатором
- 3. 3. 1. Изменение конформации РДФ-азы при взаимодействии с рибулозо-I, 5-дифосфатом (РДФ)
- 3. 3. 2. Изменение конформации молекулы РДФ-азы при взаимодействии с NaHCOg
- 3. 3. 3. Изменение конформации молекулы РДФ-азы при взаимодействии сМдС^
- 3. 4. Изменение конформации субьединиц РДФ-азы при взаимодействии с субстратом и его аналогом
- 3. 5. Избирательная модификация SH-групп РДФ-азы спин-меткой
- 3. 5. 1. Окисление нитроксильных радикалов на молекуле фермента после удаления восстановителя
- 3. 5. 2. Модификация активного центра РДФ-азы спин-меткой
- 3. 5. 3. Модификация РДФ-азы N-этилмалеинимидом с последующим спин-мечением
- 4. 1. Исследование спин-меченой РДФ-азы иммобилизованной на силикагеле, активированном глутаровым альдегидом
- 4. 2. Использование иммобилизованных препаратов РДФ-азы для поглощения COg из воздуха
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