Структурно-функциональный анализ адгезивной вариабельности пилей первого типа уропатогенных штаммов Escherichia coli
Способность адгезинов избирательно взаимодействовать с теми, или иными рецепторами, определяет тканевой тропизм патогенных вариантов кишечной палочки. Кроме того, существуют функциональные особенности адгезина FimH комменсальных штаммов, возникающие случайным образом (точечные мутации). Такие мутации в гене FimH называют патоадаптивными. Эти мутации приводят к повышению патогенного потенциала… Читать ещё >
Содержание
- Глава 1. ОБЗОР ЛИТЕРАТУРЫ
- 1. 1. Роль пилей первого типа в экологии микроорганизмов
- 1. 1. 1. Адгезии FimH как представитель класса лектинов
- 1. 1. 2. Роль пилей первого типа в адаптации микроорганизмов
- 1. 1. 3. Пили первого типа в историческом аспекте
- 1. 2. Современные представления о пилях первого типа Е. col
- 1. 2. 1. Морфология и структура пилей первого типа
- 1. 2. 2. Структурные компоненты пилей первого типа
- 1. 2. 3. Генетическая структура fim-кластера. Строение, функция 18 отдельных генов в функционировании кластера
- 1. 2. 4. Регуляция экспрессии пилей первого типа
- 1. 2. 5. Биогенез фимбрии: секреция и процессинг субъединиц, 21 сборка органеллы
- 1. 2. 6. Варианты рецепторной специфичности адгезина FimH
- 1. 3. Структурно-функциональные свойства пилей первого типа
- 1. 3. 1. Фенотипическая гетерогенность FimH штаммов Е. coli. 25 Взаимодействие с фибронектином
- 1. 3. 2. Три варианта адгезивного фенотипа FimH
- 1. 3. 3. Количественные разновидности в пределах М-фенотипа
- 1. 3. 4. Количественные различия, возникающие за счет 30 вариабельной способности связываться с одиночными маннозильными остатками
- 1. 4. Роль вариантов FimH в тканевом тропизме 31 1.4.1. Прочность фиксации на Ман-БСА коррелирует с результатами модельных опытов на уроэпителиальных клетках
- 1. 4. 2. Варианты FimH и количественные различия в связывании с ассиметричными участками мембран уроэпителия in situ
- 1. 4. 3. Варианты FimH, отличные по способности Е. coli 33 колонизировать мочевой пузырь мыши in vivo
- 1. 4. 4. Мутации FimH, повышающие уровирулентность, препятствуют адгезии к клеткам орофарингеального эпителия
- 1. 5. Варианты FimH и эволюция вирулентности
- 1. 5. 1. Филогенетический анализ аллелей FimH
- 1. 5. 2. Патоадаптивные мутации
- 1. 5. 3. Значение и перспективы дальнейшего изучения адгезина
- 1. 1. Роль пилей первого типа в экологии микроорганизмов
- 2. 1. Штаммы микроорганизмов, плазмиды и конструкции гибридов
- 2. 2. Среды, антибиотики и реактивы, используемые в работе
- 2. 2. 1. Среды
- 2. 2. 2. Антибиотики
- 2. 2. 3. Реактивы
- 2. 3. Методы исследования бакгериальной адгезии
- 2. 3. 1. Исследование агрегации дрожжей
- 2. 3. 2. Исследование агглютинации эритроцитов
- 2. 3. 3. Исследование роста (Growth assay, ИР)
- 2. 3. 4. Обогащение культуры микроорганизмов
- 2. 3. 5. Радионуклеотидное исследование адгезии
- 2. 3. 6. Исследование адгезии к клеткам защечного эпителия
- 2. 3. 7. Исследование адгезии к эритроцитам в условиях непрерывного потока
- 2. 3. 8. Исследование адгезии куроэпителиальным клеткам
- 2. 4. Иммунологические методы
- 2. 4. 1. Взаимодействие FimH антител и неимунной сыворотки со штаммами Е. coii, экспрессирующими FimH
- 2. 5. Методы работы с ДНК
- 2. 5. 1. Выделение плазмидной ДНК
- 2. 5. 2. Обработка ДНК
- 2. 6. Методы введения генетического материала в клетку
- 2. 6. 1. Классическая (кальциевая) трансформация клеток E.co
- 2. 6. 2. Электропорация
- 2. 7. Метод управляемой молекулярной динамики
- 2. 8. Математическая и статистическая обработка данных
- 3. 1. Сравнительный анализ адгезивных вариантов FimH штаммов кишечной палочки. Выявление нового адгезивного фенотипа пилей первого типа
- 3. 1. 1. Биологическая значимость структурно-функционального анализа адгезина FimH
- 3. 1. 2. FimH-FocH гибриды и их связывание с олиго- и 63 мономаннозными рецепторными субстратами
- 3. 1. 3. Чувствительность связывания с РНКазойБ и 1М-БСА к ингибированию метил-а-О-маннопиранозидом
- 3. 1. 4. Свойства гибридных FimH-FocH адгезинов при взаимодействии с эукариотическими клетками
- 3. 1. 5. Определение конфигурации рецептор-связывающего сайта
- 1. М-специфического адгезина
- 3. 1. 6. Индуцирование 1 М фенотипа FimH точечной мутацией
- 3. 1. 7. Проверка морфологии пилей первого типа и экспрессии
- 3. 1. 8. Новый адгезивный фенотип для понимания роли фенотипической гетерогенности FimH 3.2. Изучение адгезивных свойств пилей первого типа штаммов 84 Escherichia coii в условиях потока
- 3.
- 3.
- 3.
- 3.
- 3.
- 3.
Реакция агглютинации эритроцитов в статических и динамических условиях Связывание эритроцитов в камере непрерывного потока в зависимости от приложенной силы потока Моделирование конформационных изменений структуры
FimH, вызванных приложенной силой Экспериментальное подтверждение корректности управляемого компьютерного моделирования Функциональное значение адгезии, усиливаемой потоком
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