Журнал микробиологии, эпидемиологии и иммунобиологии. 2020; 97: 578-586
Протеомный анализ типичных и генетически измененных штаммов Vibrio cholerae О1 серогруппы биовара El Tor
https://doi.org/10.36233/0372-9311-2020-97-6-8Аннотация
Цель работы — сравнительное изучение экспрессии белков у типичных и генетически изменённых штаммов Vibrio cholerae O1 серогруппы биовара El Tor с помощью протеомного анализа.
Материалы и методы. В качестве модельных были использованы клинические штаммы V. cholerae — типичный M1062 (Астрахань, 1970) и генетически изменённый M1509 (Москва, 2012). Штаммы выращивали в LB бульоне рН 7,2. Получали фракции лизатов клеток и экзопротеинов и анализировали их методом 2D-электрофореза. Различающиеся белковые пятна исследовали масс-спектрометрически. Изучение выживаемости штаммов V. cholerae в условиях осмотического и оксидативного стресса проводили при инкубации штаммов в 3 М растворе NaCl или 20 мM растворе H2O2.
Результаты и обсуждение. При анализе лизатов клеток значительных отличий в экспрессии белков c известной функцией между изучаемыми штаммами не обнаружено. Подавляющая часть идентифицированных белков в лизатах функционально связана с углеводным обменом, метаболизмом аминокислот и энергетическими процессами в клетке. В то же время во фракции экзопротеинов геноварианта М1509 присутствовали в повышенных количествах белки (пероксидаза, супероксиддисмутаза, тиоредоксин, белки внешней мембраны OmpW и OmpT), защищающие клетки холерного вибриона от воздействия стрессовых факторов внешней среды. Последующее изучение устойчивости штаммов к осмотическому и оксидативному стрессу выявило лучшую выживаемость геноварианта при действии указанных факторов.
Заключение. Данные протеомного анализа типичного и генетически изменённого штаммов V. cholerae биовара El Tor свидетельствуют о повышенной экспрессии у геноварианта белков, обеспечивающих устойчивость вибрионов к действию стрессовых факторов внешней среды, что, возможно, является одной из причин их широкого распространения. Кроме того, полученные сведения позволят выявить новые биомаркеры, которые могут быть использованы для дифференциации типичных штаммов и геновариантов V. cholerae биовара El Tor.
Список литературы
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Journal of microbiology, epidemiology and immunobiology. 2020; 97: 578-586
Proteomic analysis of typical and genetically altered strains of Vibrio cholerae serogroup O1, biovar El Tor
https://doi.org/10.36233/0372-9311-2020-97-6-8Abstract
Objective — comparative study of protein expression in typical and genetically altered Vibrio cholerae strains of O1 serogroup, biovar El Tor by means of proteomic analysis.
Materials and methods. Clinical V. cholerae strains — typical strain, M106 (Astrakhan, 1970) and genetically altered one, M1509 (Moscow, 2012) — were used as model ones. Strains were cultivated in LB broth (pH7.2). Then, cell and exoprotein lysate fractions were obtained and investigated in 2D electrophoresis. Different protein stains were examined using mass spectrometry. Survivability of V. cholerae strains under osmotic and oxidative stresses was studied during incubation of the strains in 3 M NaCl solution or 20 mM H2O2 solution.
Results and discussion. When analyzing cell lysates, significant differences in protein expression with known function between studied strains were not detected. The great majority of identified proteins in the lysates is functionally associated with carbohydrate metabolism, amino acid metabolism, and energy processes in a cell. At the same time, exoprotein fraction of M1509 genovariant contained increased amount of proteins (peroxidase, superoxide dismutase, thioredoxin, outer membrane proteins OmpW, OmpT) protecting the cells of cholera vibrio from effect of stress factors of the environment. Further study of the resistance to osmotic and oxidative stresses revealed better survivability in the genovariant when exposed to the stated factors.
Conclusion. The data of proteomic analysis of the typical and genetically altered V. cholera strains, biovar El Tor, testify to high levels of expression of the proteins that provide for vibrio resistance to the effect of environmental stress factors in genovariants, which is possibly one of the causes of their wide dissemination. In addition, the results obtained will allow for identification of new biomarkers which can be used for differentiation of typical strains and genovariants of V. cholerae, biovar El Tor.
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