Скорострельная протеомика в изучении адаптивных реакций растений на стресс

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Аннотация

В связи со стремительным изменением климата засуха, экстремально высокие температуры и засоление почв стали заметными экологическими стрессорами, которые на данный момент встречаются в большинстве наземных экосистем. В связи с исключительным влиянием этих факторов на сельское хозяйство, биохимия и молекулярная биология стрессовых реакций растений последние несколько десятилетий остается в центре исследовательского интереса во всем мире. Так, протеомика «снизу-вверх» стала универсальным инструментом исследования растений в целом и биологии клеточного стресса в частности. Растения считаются сложным объектом для исследования: их клетки богаты полисахаридами, полифенолами и труднорастворимыми в воде белками, поэтому их протеом обычно анализируют с помощью электрофоретических методов. Однако последние достижения в области методов пробоподготовки (в первую очередь, солюбилизации и переваривания белков) позволили создать «безгелевые» методы для образцов растительного происхождения. Внедрение высокопроизводительной нанопоточной обратно-фазовой высокоэффективной жидкостной хроматографии, соединенной с масс-спектрометрией с ионизацией электрораспылением (nanoRP-HPLC-ESI-MS), обеспечило доступ к массивам данных с высокой скоростью идентификации белков. Кроме того, высокая воспроизводимость высокоэффективной жидкостной хроматографии позволяет проводить высокочувствительное и точное количественное определение. Поэтому в последнее десятилетие дробовая протеомика стала методом выбора при изучении адаптивных стрессовых реакций протеома растений. В данном обзоре рассматриваются стратегии шотган-протеомики «снизу-вверх» в биологии растений и обсуждается ее применение для изучения стрессовых реакций растений. В статье также обсуждаются основные этапы анализа протеома растений, возникающие проблемы и перспективы.

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Об авторах

Дарья Петровна Горбач

Институт физиологии растений им. К.А. Тимирязева

Email: daria.gorba4@yandex.ru
ORCID iD: 0000-0001-6836-5004
Россия, Москва

Татьяна Сергеевна Леонова

Институт биохимии растений им. Лейбница

Email: tatiana.leonova@ipb-halle.de
ORCID iD: 0000-0002-7153-5059
SPIN-код: 6132-3216
Германия, Галле (Заале)

Анастасия Андреевна Орлова

Институт физиологии растений им. К.А. Тимирязева

Email: lanas_95@mail.ru
ORCID iD: 0000-0002-7836-5785

канд. фарм. наук

Россия, Москва

Надежда Владимировна Фролова

Институт физиологии растений им. К.А. Тимирязева

Email: frolovanadja@yandex.ru
ORCID iD: 0000-0002-1895-1133
SPIN-код: 1633-7805

канд. биол. наук

Россия, Москва

Анастасия Кирилловна Гурина

Институт физиологии растений им. К.А. Тимирязева; Санкт-Петербургский государственный университет

Email: gnastyak0@gmail.com
ORCID iD: 0009-0000-6514-1240
Россия, Москва; Санкт-Петербург

Катерина Владимировна Данько

Санкт-Петербургский государственный университет

Email: danko_katerina@mail.ru
ORCID iD: 0000-0003-3987-2175
Россия, Санкт-Петербург

Елена Михайловна Лукашева

Санкт-Петербургский государственный университет

Email: elena_lukasheva@mail.ru
ORCID iD: 0000-0002-0889-2395
SPIN-код: 6071-3591
Россия, Санкт-Петербург

Мария Александровна Черевацкая

Санкт-Петербургский государственный университет

Email: maria.cherevatskaya@gmail.com
ORCID iD: 0000-0002-5604-3400
SPIN-код: 9395-1519

канд. хим. наук

Россия, Санкт-Петербург

Елена Викторовна Цветкова

Санкт-Петербургский государственный университет

Email: evtsvetkova72@mail.ru
ORCID iD: 0000-0002-2022-615X
SPIN-код: 9221-5326

канд. биол. наук

Россия, Санкт-Петербург

Андрей Александрович Фролов

Институт физиологии растений им. К.А. Тимирязева

Автор, ответственный за переписку.
Email: Andrej.Frolov@ipb-halle.de
ORCID iD: 0000-0003-3250-5858
SPIN-код: 5105-2490

д-р биол. наук

Россия, Москва

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2. Рис. 1. Эффекты, оказываемые активными формами кислорода (АФК) в присутствии внешних факторов стрессора разной амплитуды.

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3. Рис. 2. Пути образования активных форм кислорода (ROS) в растительной клетке. O2– — супероксидный радикал; H2O2 — перекись водорода; 1O2 — синглетный кислород; OH– — гидроксильный радикал; PSI — фотосистема I; SOD — супероксиддисмутаза; APX — аскорбатпероксидаза; AA — аскорбиновая кислота; MDA — малоновый диальдегид; NADH — никотинамидадениндинуклеотид; MDHAR — монодегидроаскорбатредуктаза; DHA — докозагексаеновая кислота; GSSG — глутатион дисульфид; GSH — глутатион; NADP — никотинамидадениндинуклеотидфосфат; КАТ — каталаза; GPX — глутатион пероксидаза.

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4. Рис. 3. Схема передачи биологической информации в клетке.

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5. Рис. 4. Две стратегии шотган масс-спектрометрии: методы «сверху вниз» и «снизу вверх».

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