Новые подходы в изучении структурной и функциональной организации систем анионного транспорта в биологических мембранах
При изучении механизмов и энергетических источников транспорта различных веществ через мембраны эпителиальных клеток использовались разные методические подходы, равно как и разные объекты исследования. Среди них отметим эксперименты in situ с использованием микроперфузии просвета проксимального канальца или периферийной кровяной капиллярной сети (Sheridan, et. al., 1983; Fritzsch, et. al., 1984… Читать ещё >
Содержание
- Список принятых сокращений
Глава 1. Топография транспортных систем проксимальных канальцев почки и структурная полярность эпителиальных клеток.
Глава 2. Транспорт органических анионов в проксимальных канальцах почки.
2.1. Локализация.
2.2. Специфичность.
2.3. Этапы секреции органических анионов в почечных канальцах.
2.3.1. Транспорт через базолатеральную мембрану.
2.3.2. Внутриклеточное распределение органических анионов.
2.3.3. Транспорт через апикальную мембрану.
Глава 3. Выделение апикальной и базолатеральной мембран из клеток проксимальных канальцев и возможность изучения транспорта органических анионов на везикулярных структурах.
Глава 4. Влияние фазового состояния липидного бислоя на функционирование мембранных белков.
Глава 5. Материалы и методы исследования транспорта органических анионов в проксимальных канальцах почки.
5.1. Выделение везикулярных препаратов апикальной и базолатеральной мембраны из коры почки крысы.
5.2. Определение активности маркерных ферментов.
5.3. Обсуждение метода выделения мембранных фракций.
5.4. Определение уровня накопления [3Н]-п-аминогиппуровой кислоты ([3Н]ПАГ) в везикулярных мембранных препаратах.
5.5. Выбор оптимальных условий фильтрации везикулярных мембранных препаратов.
5.6. Метод введения ЭПР зонда в липидный бислой AM и математическая обработка данных ЭПР спектроскопии.
5.7. Электронная микроскопия фракции AM.
Глава 6. Результаты исследования транспорта органических анионов с использованием везикулярных препаратов апикальной мембраны. Влияние фазового состояния липидного бислоя на транспортную кинетику.
6.1. Электронно-микроскопический контроль фракции апикальной мембраны.
6.2. Изучение транспорта органических анионов в везикулы АМ коры почки крысы.
Глава 7. Обсуждение результатов исследования транспорта органических анионов через апикальную мембрану проксимальных канальцев почки крысы.
7.1. Общая характеристика накопления ПАГ в везикулах апикальной мембраны.
7.2. Влияние фазового состояния липидного бислоя АМ на кинетические характеристики переносчика ПАГ и активность щелочной фосфатазы.
Глава 8. Идентификация переносчиков органических анионов проксимальных канальцев почки.
8.1. Молекулярно-биологическое клонирование и метод аффинных меток.
8.2. Материалы и методы.
8.2.1. Синтез бромацетилированной п-аминогиппуровой кислоты.
8.2.2. Синтез диазогиппуровой кислоты.
8.2.3. Обработка мембранной фракции бромацетилированным п-аминогиппуратом.
8.2.4. Обработка мембранной фракции диазогиппуратом.
8.2.5. Разделение белковой и липидной фракции перед электрофорезом и электрофорез.
8.2.6. Выделение и анализ липидов.
8.3. Необратимое ингибирование транспорта ПАГ в везикулах апикальной мембраны бромацетилированным-п-аминогиппуратом (ВгАсПАГ).
8.4. Аффинная идентификация полипептидов системы транспорта органических анионов апикальной и базолатеральной мембраны проксимальных канальцев почки.
Глава 9. Математическое моделирование транспортных потоков в биологических мембранах.
9.1. Кинетика и механизм транспортной реакции. История развития.
9.2. Основные черты транспортной реакции. Современные представления.
9.2.1. Классификация транспортных процессов.
9.2.2. Вторично-активный транспорт.
9.3. Способы интерпретации транспортных (векторных) реакций.
9.4. Противопоток.
9.5. Котранспорт.
Глава 10. Экспериментальные примеры функционального взаимодействия котранспортных и анион-обменных переносчиков.
10.1. Транспортная система обменного типа мочевая кислота (урат)/ анион.
10.2. Транспортная система обменного типа монокарбоксилат/ОН.
10.3. Cl/формат переносчик обменного типа.
10.4. С1/оксалат переносчик обменного типа.
10.5. S0427HCCh переносчик обменного типа.
10.6. Na -независимый СГ/ НСОз переносчик обменного типа.
10.7. Анионообменные переносчики в других тканях.
Глава 11. Минимальная математическая модель энергетического сопряжения котранспортного и анион-обменного механизмов переноса в биологических мембранах.
11.1. Исходные предположения и формулировка модели.
11.2. Компьютерное моделирование кооперативной работы котранспортного и анион-обменного переносчиков.
11.3. Использование математической модели для описания экспериментальных результатов.
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