Paulina Patalas-Krawczyk

Instytut Biologii Doświadczalnej im. Marcelego Nenckiego

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Jerzy Duszyński

Instytut Biologii Doświadczalnej im. Marcelego Nenckiego

Jędrzej Szymański

Instytut Biologii Doświadczalnej im. Marcelego Nenckiego

Bernadeta Michalska

Instytut Biologii Doświadczalnej im. Marcelego Nenckiego

Mariusz R. Więckowski

Instytut Biologii Doświadczalnej im. Marcelego Nenckiego

Joanna Szczepanowska

Instytut Biologii Doświadczalnej im. Marcelego Nenckiego

Robert Hołyst

Institute of Physical Chemistry

Tomasz Kalwarczyk

Institute of Physical Chemistry

Krzysztof Szczepański

Polish Academy of Sciences

Dominika Malińska

Instytut Biologii Doświadczalnej im. Marcelego Nenckiego

Karina Kwapiszewska

Polish Academy of Sciences

Cooperative Institutions

Polish Academy of Sciences

Instytut Biologii Doświadczalnej im. Marcelego Nenckiego

Poznan University of Medical Sciences

University of Warsaw

Philip Morris International (Switzerland)

Institute of Physical Chemistry

University of Ferrara

Wojskowy Instytut Medycyny Lotniczej

University of Pittsburgh

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Interaction of Mitochondria with the Endoplasmic Reticulum and Plasma Membrane in Calcium Homeostasis, Lipid Trafficking and Mitochondrial Structure

International Journal of Molecular Sciences (2017)

Jędrzej Szymański Justyna Janikiewicz Bernadeta Michalska Paulina Patalas-Krawczyk Mariasole Perrone

Studying organelles in isolation has been proven to be indispensable for deciphering the underlying mechanisms of molecular cell biology. However, observing organelles in intact cells with the use of microscopic techniques reveals a new set of different junctions and contact sites between them that contribute to the control and regulation of various cellular processes, such as calcium and lipid exchange or structural reorganization of the mitochondrial network. In recent years, many studies focused their attention on the structure and function of contacts between mitochondria and other organelles. From these studies, findings emerged showing that these contacts are involved in various processes, such as lipid synthesis and trafficking, modulation of mitochondrial morphology, endoplasmic reticulum (ER) stress, apoptosis, autophagy, inflammation and Ca 2 + handling. In this review, we focused on the physical interactions of mitochondria with the endoplasmic reticulum and plasma membrane and summarized present knowledge regarding the role of mitochondria-associated membranes in calcium homeostasis and lipid metabolism.

Membrane contact site

Organelle

Calcium Signaling

Lipid droplet

Homeostasis

10.3390/ijms18071576

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Citations (200)

Exosomes in Plasma of Patients with Ovarian Carcinoma: Potential Biomarkers of Tumor Progression and Response to Therapy

Gynecology & Obstetrics (2012)

Marta Szajnik Magdalena Derbis Michał Lach Paulina Patalas-Krawczyk Marcin Michalak

In patients with Ovarian Cancer (OvCa) exosomes released by tumor cells are present in the plasma and could be involved in tumor progression. This study examines the association between the exosome presence/protein content in plasma of OvCa patients and disease outcome, response to standard therapy and/or tumorresistance to therapies in patients studied at diagnosis and also serially during and after therapy.Exosomes were purified from OvCa patients' plasma (n=22), patients with benign tumors (n=10) or (n=10) healthy controls (NC) using ultracentrifugation. Exosomes were visualized by scanning electron microscopy. Their protein content was measured. The presence of MAGE 3/6 and TGF-β1 in exosomes was evaluated in Western blots.The OvCa patients' plasma contained higher levels of exosomal proteins (p<0.05) compared to those isolated from plasma of patients with benign tumors or NC. Exosomes isolated from OvCa patients's plasma carried TGF-β1 and MAGE3/6, which distinguished OvCa patients from those with benign tumors and NC. High protein levels of exosomes were seen in newly diagnosed patients; however in advanced stages of OvCa patients the protein content of isolated exosomes was significantly higher than that of early stages. The exosome levels variably changed during/after chemotherapy, and correlations between the changes in exosomal protein levels and clinical data suggested that the protein content of exosomes might be useful in predicting responses to therapy and prognosis in OvCa patients.Analysis of plasma exosomes levels offers a novel approach to diagnosis and monitoring response to therapies in OvCa patients.

Exosome

10.4172/2161-0932.s4-003

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Apparent Anomalous Diffusion in the Cytoplasm of Human Cells: The Effect of Probes’ Polydispersity

The Journal of Physical Chemistry B (2017)

Tomasz Kalwarczyk Karina Kwapiszewska Krzysztof Szczepański Krzysztof Sozański Jędrzej Szymański

This work, based on in vivo and in vitro measurements, as well as in silico simulations, provides a consistent analysis of diffusion of polydisperse nanoparticles in the cytoplasm of living cells. Using the example of fluorescence correlation spectroscopy (FCS), we show the effect of polydispersity of probes on the experimental results. Although individual probes undergo normal diffusion, in the ensemble of probes, an effective broadening of the distribution of diffusion times occurs—similar to anomalous diffusion. We introduced fluorescently labeled dextrans into the cytoplasm of HeLa cells and found that cytoplasmic hydrodynamic drag, exponentially dependent on probe size, extraordinarily broadens the distribution of diffusion times across the focal volume. As a result, the in vivo FCS data were effectively fitted with the anomalous subdiffusion model while for a monodisperse probe the normal diffusion model was most suitable. Diffusion time obtained from the anomalous diffusion model corresponds to a probe whose size is determined by the weight-average molecular weight of the polymer. The apparent anomaly exponent decreases with increasing polydispersity of the probes. Our results and methodology can be applied in intracellular studies of the mobility of nanoparticles, polymers, or oligomerizing proteins.

Dispersity

Anomalous Diffusion

Fluorescence Correlation Spectroscopy

10.1021/acs.jpcb.7b07158

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Citations (46)

Mitochondria as a possible target for nicotine action

Journal of Bioenergetics and Biomembranes (2019)

Dominika Malińska Mariusz R. Więckowski Bernadeta Michalska Karolina Drabik Monika Prill

Mitochondria are multifunctional and dynamic organelles deeply integrated into cellular physiology and metabolism. Disturbances in mitochondrial function are involved in several disorders such as neurodegeneration, cardiovascular diseases, metabolic diseases, and also in the aging process. Nicotine is a natural alkaloid present in the tobacco plant which has been well studied as a constituent of cigarette smoke. It has also been reported to influence mitochondrial function both in vitro and in vivo. This review presents a comprehensive overview of the present knowledge of nicotine action on mitochondrial function. Observed effects of nicotine exposure on the mitochondrial respiratory chain, oxidative stress, calcium homeostasis, mitochondrial dynamics, biogenesis, and mitophagy are discussed, considering the context of the experimental design. The potential action of nicotine on cellular adaptation and cell survival is also examined through its interaction with mitochondria. Although a large number of studies have demonstrated the impact of nicotine on various mitochondrial activities, elucidating its mechanism of action requires further investigation.

10.1007/s10863-019-09800-z

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Citations (83)

Effects of plant alkaloids on mitochondrial bioenergetic parameters

Food and Chemical Toxicology (2021)

Paulina Patalas-Krawczyk Dominika Malińska Jarosław Walczak Gilles Kratzer Monika Prill

Mitochondria are among the first responders to various stress factors that challenge cell and tissue homeostasis. Various plant alkaloids have been investigated for their capacity to modulate mitochondrial activities. In this study, we used isolated mitochondria from mouse brain and liver tissues to assess nicotine, anatabine and anabasine, three alkaloids found in tobacco plant, for potential modulatory activity on mitochondrial bioenergetics parameters. All alkaloids decreased basal oxygen consumption of mouse brain mitochondria in a dose-dependent manner without any effect on the ADP-stimulated respiration. None of the alkaloids, at 1 nM or 1.25 μM concentrations, influenced the maximal rate of swelling of brain mitochondria. In contrast to brain mitochondria, 1.25 μM anatabine, anabasine and nicotine increased maximal rate of swelling of liver mitochondria suggesting a toxic effect. Only at 1 mM concentration, anatabine slowed down the maximal rate of Ca2+-induced swelling and increased the time needed to reach the maximal rate of swelling. The observed mitochondrial bioenergetic effects are probably mediated through a pathway independent of nicotinic acetylcholine receptors, as quantitative proteomic analysis could not confirm their expression in pure mitochondrial fractions isolated from mouse brain tissue.

Anabasine

Bioenergetics

Coenzyme Q10

10.1016/j.fct.2021.112316

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Mitochondria and Reactive Oxygen Species in Aging and Age-Related Diseases

International review of cell and molecular biology (2018)

Carlotta Giorgi Saverio Marchi Inês C. M. Simões Ziyu Ren Giampaolo Morciano

Cell Physiology

Organelle

10.1016/bs.ircmb.2018.05.006

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Citations (303)

Determination of oligomerization state of Drp1 protein in living cells at nanomolar concentrations

Scientific Reports (2019)

Karina Kwapiszewska Tomasz Kalwarczyk Bernadeta Michalska Krzysztof Szczepański Jędrzej Szymański

Biochemistry in living cells is an emerging field of science. Current quantitative bioassays are performed ex vivo, thus equilibrium constants and reaction rates of reactions occurring in human cells are still unknown. To address this issue, we present a non-invasive method to quantitatively characterize interactions (equilibrium constants, KD) directly within the cytosol of living cells. We reveal that cytosolic hydrodynamic drag depends exponentially on a probe's size, and provide a model for its determination for different protein sizes (1-70 nm). We analysed oligomerization of dynamin-related protein 1 (Drp1, wild type and mutants: K668E, G363D, C505A) in HeLa cells. We detected the coexistence of wt-Drp1 dimers and tetramers in cytosol, and determined that KD for tetramers was 0.7 ± 0.5 μM. Drp1 kinetics was modelled by independent simulations, giving computational results which matched experimental data. This robust method can be applied to in vivo determination of KD for other protein-protein complexes, or drug-target interactions.

10.1038/s41598-019-42418-0

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Nanoscale Viscosity of Cytoplasm Is Conserved in Human Cell Lines

The Journal of Physical Chemistry Letters (2020)

Karina Kwapiszewska Krzysztof Szczepański Tomasz Kalwarczyk Bernadeta Michalska Paulina Patalas-Krawczyk

Metabolic reactions in living cells are limited by diffusion of reagents in the cytoplasm. Any attempt to quantify the kinetics of biochemical reactions in the cytosol should be preceded by careful measurements of the physical properties of the cellular interior. The cytoplasm is a complex, crowded fluid characterized by effective viscosity dependent on its structure at a nanoscopic length scale. In this work, we present and validate the model describing the cytoplasmic nanoviscosity, based on measurements in seven human cell lines, for nanoprobes ranging in diameters from 1 to 150 nm. Irrespective of cell line origin (epithelial–mesenchymal, cancerous–noncancerous, male–female, young–adult), we obtained a similar dependence of the viscosity on the size of the nanoprobes, with characteristic length-scales of 20 ± 11 nm (hydrodynamic radii of major crowders in the cytoplasm) and 4.6 ± 0.7 nm (radii of intercrowder gaps). Moreover, we revealed that the cytoplasm behaves as a liquid for length scales smaller than 100 nm and as a physical gel for larger length scales.

Length scale

10.1021/acs.jpclett.0c01748

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Citations (72)

Mitochondria-associated membranes in aging and senescence: structure, function, and dynamics

Cell Death and Disease (2018)

Justyna Janikiewicz Jędrzej Szymański Dominika Malińska Paulina Patalas-Krawczyk Bernadeta Michalska

Sites of close contact between mitochondria and the endoplasmic reticulum (ER) are known as mitochondria-associated membranes (MAM) or mitochondria-ER contacts (MERCs), and play an important role in both cell physiology and pathology. A growing body of evidence indicates that changes observed in the molecular composition of MAM and in the number of MERCs predisposes MAM to be considered a dynamic structure. Its involvement in processes such as lipid biosynthesis and trafficking, calcium homeostasis, reactive oxygen species production, and autophagy has been experimentally confirmed. Recently, MAM have also been studied in the context of different pathologies, including Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, type 2 diabetes mellitus and GM1-gangliosidosis. An underappreciated amount of data links MAM with aging or senescence processes. In the present review, we summarize the current knowledge of basic MAM biology, composition and action, and discuss the potential connections supporting the idea that MAM are significant players in longevity.

Senescence

10.1038/s41419-017-0105-5

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Citations (180)

Insight into the fission mechanism by quantitative characterization of Drp1 protein distribution in the living cell

Scientific Reports (2018)

Bernadeta Michalska Karina Kwapiszewska Joanna Szczepanowska Tomasz Kalwarczyk Paulina Patalas-Krawczyk

One of the main players in the process of mitochondrial fragmentation is dynamin-related protein 1 (Drp1), which assembles into a helical ring-like structure on the mitochondria and facilitates fission. The fission mechanism is still poorly understood and detailed information concerning oligomeric form of Drp1, its cellular distribution and the size of the fission complex is missing. To estimate oligomeric forms of Drp1 in the cytoplasm and on the mitochondria, we performed a quantitative analysis of Drp1 diffusion and distribution in gene-edited HeLa cell lines. This paper provides an insight into the fission mechanism based on the quantitative description of Drp1 cellular distribution. We found that approximately half of the endogenous GFP-Drp1 pool remained in the cytoplasm, predominantly in a tetrameric form, at a concentration of 28 ± 9 nM. The Drp1 mitochondrial pool included many different oligomeric states with equilibrium distributions that could be described by isodesmic supramolecular polymerization with a Kd of 31 ± 10 nM. We estimated the average number of Drp1 molecules forming the functional fission complex to be approximately 100, representing not more than 14% of all Drp1 oligomers. We showed that the upregulated fission induced by niclosamide is accompanied by an increase in the number of large Drp1 oligomers.

10.1038/s41598-018-26578-z

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Citations (40)