Злобовская Ольга Анатольевна

Образование

Период обученияСтрана, городУчебное заведениеДополнительная информация
2012 Россия, Москва Институт биоорганической химии им. академиков М.М. Шемякина и Ю.А. Овчинникова
1997–2007 Россия, Москва гимназия №1543 золотая медаль, дипломы с олимпиад по физике, химии, биологии
2007–2012 Россия, Москва МГУ им. М.В. Ломоносова премия имени Р.Б. Хесина (2011 г.)

Научные интересы

Молекулярная биология, фототоксичные флуоресцентные белки, апоптоз, реал-тайм ПЦР

Основные научные результаты

Дальнекрасный сенсор на активность каспазы 3 (патент)

Гранты и проекты

ПериодДополнительная информация
РФФИ 14-04-31600-мол_а (ЦИТиС 01201452918).«Генетически кодируемые инфракрасные флуоресцентные сенсоры ионов кальция и пероксида водорода».Руководитель - Саркисян К.С.
Программа Президиума РАН «Молекулярная и клеточная биология».Тема «Оптогенетические подходы к визуализации тонкой клеточной организации и направленному вмешательству в клеточные процессы».Руководитель - д.б.н. Лукьянов К.А.
Программа Президиума РАН «Фундаментальные исследований для разработки медицинских технологий».Тема «Разработка технологии преодоления иммунологической толерогенности опухоли с помощью ключевого модулятора активности иммунокомпетентных клеток белка OX40L».Руководитель - Лукьянов С.А.
РНФ 14-25-00129«Разработка новейших подходов для изучения механизмов действия противоопухолевых препаратов и раннего ответа опухоли на лечение на основе оптических и молекулярных технологий». Руководитель - д.б.н. Лукьянов К.А.
Грант РНФ 14-35-00105"Комплексное исследование молекулярной эволюции злокачественных опухолей для разработки персонифицированных подходов к ведению онкологических больных"Руководитель - Лукьянов С.А.
РФФИ 14-04-01185 А (ЦИТиС 01201452921)Экспрессия OX40L на поверхности клеток перевиваемых опухолей как инструмент для индукции эффективного противоопухолевого ответа.Руководитель - к.б.н. Серебровская Е.О.

Избранные публикации

  1. Sergeeva T.F., Shirmanova M.V., Zlobovskaya O.A., Gavrina A.I., Dudenkova V.V., Lukina M.M., Lukyanov K.A., Zagaynova E.V. (2017). Relationship between intracellular pH, metabolic co-factors and caspase-3 activation in cancer cells during apoptosis. Biochim. Biophys. Acta 1864 (3), 604–611 [+]

    A complex cascade of molecular events occurs in apoptotic cells but cell-to-cell variability significantly complicates determination of the order and interconnections between different processes. For better understanding of the mechanisms of programmed cell death, dynamic simultaneous registration of several parameters is required. In this paper we used multiparameter fluorescence microscopy to analyze energy metabolism, intracellular pH and caspase-3 activation in living cancer cells in vitro during staurosporine-induced apoptosis. We performed metabolic imaging of two co-factors, NAD(P)H and FAD, and used the genetically encoded pH-indicator SypHer1 and the FRET-based sensor for caspase-3 activity, mKate2-DEVD-iRFP, to visualize these parameters by confocal fluorescence microscopy and two-photon fluorescence lifetime imaging microscopy. The correlation between energy metabolism, intracellular pH and caspase-3 activation and their dynamic changes were studied in CT26 cancer cells during apoptosis. Induction of apoptosis was accompanied by a switch to oxidative phosphorylation, cytosol acidification and caspase-3 activation. We showed that alterations in cytosolic pH and the activation of oxidative phosphorylation are relatively early events associated with the induction of apoptosis.

    ID:1709
  2. Ryumina A.P., Serebrovskaya E.O., Staroverov D.B., Zlobovskaya O.A., Shcheglov A.S., Lukyanov S.A., Lukyanov K.A. (2016). Lysosome-associated miniSOG as a photosensitizer for mammalian cells. BioTechniques 61 (2), 92–4 [+]

    Genetically encoded photosensitizers represent a promising optogenetic tool for the induction of light-controlled oxidative stress strictly localized to a selected intracellular compartment. Here we tested the phototoxic effects of the flavin-containing phototoxic protein miniSOG targeted to the cytoplasmic surfaces of late endosomes and lysosomes by fusion with Rab7. In HeLa Kyoto cells stably expressing miniSOG-Rab7, we demonstrated a high level of cell death upon blue-light illumination. Pepstatin A completely abolished phototoxicity of miniSOG-Rab7, showing a key role for cathepsin D in this model. Using a far-red fluorescence sensor for caspase-3, we observed caspase-3 activation during miniSOG-Rab7-mediated cell death. We conclude that upon illumination, miniSOG-Rab7 induces lysosomal membrane permeabilization (LMP) and leakage of cathepsins into the cytosol, resulting in caspase-dependent apoptosis.

    ID:1555
  3. Zlobovskaya O.A., Sergeeva T.F., Shirmanova M.V., Dudenkova V.V., Sharonov G.V., Zagaynova E.V., Lukyanov K.A. (2016). Genetically encoded far-red fluorescent sensors for caspase-3 activity. BioTechniques 60 (2), 62–8 [+]

    Caspase-3 is a key effector caspase that is activated in both extrinsic and intrinsic pathways of apoptosis. Available fluorescent sensors for caspase-3 activity operate in relatively short wavelength regions and are nonoptimal for multiparameter microscopy and whole-body imaging. In the present work, we developed new genetically encoded sensors for caspase-3 activity possessing the most red-shifted spectra to date. These consist of Förster resonance energy transfer (FRET) pairs in which a far-red fluorescent protein (mKate2 or eqFP650) is connected to the infrared fluorescent protein iRFP through a linker containing the DEVD caspase-3 cleavage site. During staurosporine-induced apoptosis of mammalian cells (HeLa and CT26), both mKate2-DEVD-iRFP and eqFP650-DEVD-iRFP sensors showed a robust response (1.6-fold increase of the donor fluorescence intensity). However, eqFP650-DEVD-iRFP displayed aggregation in some cells. For stably transfected CT26 mKate2-DEVD-iRFP cells, fluorescence lifetime imaging (FLIM) enabled us to detect caspase-3 activation due to the increase of mKate2 donor fluorescence lifetime from 1.45 to 2.05 ns. We took advantage of the strongly red-shifted spectrum of mKate2-DEVD-iRFP to perform simultaneous imaging of EGFP-Bax translocation during apoptosis. We conclude that mKate2-DEVD-iRFP is well-suited for multiparameter imaging and also potentially beneficial for in vivo imaging in animal tissues.

    ID:1373
  4. Zlobovskaya O.A., Sarkisyan K.S., Lukyanov K.A. (2015). Infrared Fluorescent Protein iRFP as an Acceptor for Förster Resonance Energy Transfer. Bioorg. Khim. 41 (3), 299–304 [+]

    Bacteriophytochrome-based infrared fluorescent protein iRFP was tested as an acceptor for F6rster resonance energy transfer (FRET). Far-red GFP-like fluorescent proteins mKate2, eqFP650, and eqFP670 were used as donors; Bacterial expression vectors encoding donor and acceptor proteins fused by a 17-amino acid linker were.constructed. FRET for purified proteins in vitro was, estimated from increase of the donor emission after digestion of the linker. Among the three constructs tested, the most efficient FRET (approximately 30%) was detected for the eqFP650-iRFP pair.

    ID:1329
  5. Sarkisyan K.S., Zlobovskaya O.A., Gorbachev D.A., Bozhanova N.G., Sharonov G.V., Staroverov D.B., Egorov E.S., Ryabova A.V., Solntsev K.M., Mishin A.S., Lukyanov K.A. (2015). KillerOrange, a Genetically Encoded Photosensitizer Activated by Blue and Green Light. PLoS ONE 10 (12), e0145287 [+]

    Genetically encoded photosensitizers, proteins that produce reactive oxygen species when illuminated with visible light, are increasingly used as optogenetic tools. Their applications range from ablation of specific cell populations to precise optical inactivation of cellular proteins. Here, we report an orange mutant of red fluorescent protein KillerRed that becomes toxic when illuminated with blue or green light. This new protein, KillerOrange, carries a tryptophan-based chromophore that is novel for photosensitizers. We show that KillerOrange can be used simultaneously and independently from KillerRed in both bacterial and mammalian cells offering chromatic orthogonality for light-activated toxicity.

    ID:1355