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Article ; Online: Malondialdehyde enhances PsbP protein release during heat stress in Arabidopsis.

Kumar, Aditya / Prasad, Ankush / Sedlářová, Michaela / Pospíšil, Pavel

2023 Volume 202, Page(s) 107984

Abstract: Under environmental conditions, plants are exposed to various abiotic and biotic stress factors, which commonly cause the oxidation of lipids and proteins. Lipid peroxidation constantly produces malondialdehyde (MDA), a secondary product of lipid ... ...

Abstract	Under environmental conditions, plants are exposed to various abiotic and biotic stress factors, which commonly cause the oxidation of lipids and proteins. Lipid peroxidation constantly produces malondialdehyde (MDA), a secondary product of lipid peroxidation, which is covalently bound to proteins forming MDA-protein adducts. The spatial distribution of MDA-protein adducts in Arabidopsis leaves shows that MDA-protein adducts are located in the chloroplasts, uniformly spread out over the thylakoid membrane. At the lumenal side of thylakoid membrane, MDA interacts with PsbP, an extrinsic subunit of the photosystem II (PSII), which is in electrostatic interaction with the PSII core proteins. Under heat stress, when MDA is moderately enhanced, the electrostatic interaction between PsbP and PSII core proteins is weakened, and PsbP with bound MDA is released in the lumen. It is proposed here that the electrophilic MDA is bound to the nucleophilic lysine residues of PsbP, which are involved in electrostatic interactions with the negatively charged glutamate of the PSII core protein. Our data provide crucial information about the MDA binding topology in the higher plant PSII complex, which is necessary to understand better the physiological functions of MDA for plant survival under stress.
MeSH term(s)	Malondialdehyde ; Arabidopsis ; Glutamic Acid ; Lysine ; Heat-Shock Response
Chemical Substances	Malondialdehyde (4Y8F71G49Q) ; Glutamic Acid (3KX376GY7L) ; Lysine (K3Z4F929H6)
Language	English
Publishing date	2023-08-23
Publishing country	France
Document type	Journal Article
ZDB-ID	742978-2
ISSN	1873-2690 ; 0981-9428
ISSN (online)	1873-2690
ISSN	0981-9428
DOI	10.1016/j.plaphy.2023.107984
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Article ; Online: Oxidative modification of collagen by malondialdehyde in porcine skin.

Paculová, Vendula / Prasad, Ankush / Sedlářová, Michaela / Pospíšil, Pavel

Archives of biochemistry and biophysics

2023 Volume 752, Page(s) 109850

Abstract: Human skin is exposed to various physical and chemical stress factors, which commonly cause the oxidation of lipids and proteins. In this study, azo initiator AAPH [2,2' -azobis(2-methylpropionamidine) dihydrochloride] was employed to initiate lipid ... ...

Abstract	Human skin is exposed to various physical and chemical stress factors, which commonly cause the oxidation of lipids and proteins. In this study, azo initiator AAPH [2,2' -azobis(2-methylpropionamidine) dihydrochloride] was employed to initiate lipid peroxidation in porcine skin as an ex vivo model for human skin. We demonstrate that malondialdehyde (MDA), a secondary product of lipid peroxidation, is covalently bound to collagen in the dermis, forming MDA-collagen adducts. The binding of MDA to collagen results in an unfolding of the collagen triple helix, formation of the dimer of α-chains of collagen, and fragmentation of the collagen α-chain. It is proposed here that the MDA is bound to the lysine residues of α-chain collagen, which are involved in electrostatic interaction and hydrogen bonding with the glutamate and aspartate of other α-chains of the triple helix. Our data provide crucial information about the MDA binding topology in the skin, which is necessary to understand better the various types of skin-related diseases and the aging process in the skin under stress.
MeSH term(s)	Humans ; Swine ; Malondialdehyde/metabolism ; Oxidation-Reduction ; Collagen/metabolism ; Lipid Peroxidation ; Oxidative Stress ; Animals
Chemical Substances	Malondialdehyde (4Y8F71G49Q) ; Collagen (9007-34-5)
Language	English
Publishing date	2023-12-06
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	523-x
ISSN	1096-0384 ; 0003-9861
ISSN (online)	1096-0384
ISSN	0003-9861
DOI	10.1016/j.abb.2023.109850
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Article ; Online: Detection and characterization of free oxygen radicals induced protein adduct formation in differentiating macrophages.

Manoharan, Renuka Ramalingam / Sedlářová, Michaela / Pospíšil, Pavel / Prasad, Ankush

Biochimica et biophysica acta. General subjects

2023 Volume 1867, Issue 5, Page(s) 130324

Abstract: Reactive oxygen species play a key role in cellular homeostasis and redox signaling at physiological levels, where excessive production affects the function and integrity of macromolecules, specifically proteins. Therefore, it is important to define ... ...

Abstract	Reactive oxygen species play a key role in cellular homeostasis and redox signaling at physiological levels, where excessive production affects the function and integrity of macromolecules, specifically proteins. Therefore, it is important to define radical-mediated proteotoxic stress in macrophages and identify target protein to prevent tissue dysfunction. A well employed, THP-1 cell line was utilized as in vitro model to study immune response and herein we employ immuno-spin trapping technique to investigate radical-mediated protein oxidation in macrophages. Hydroxyl radical formation along macrophage differentiation was confirmed by electron paramagnetic resonance along with confocal laser scanning microscopy using hydroxyphenyl fluorescein. Lipid peroxidation product, malondialdehyde, generated under experimental conditions as detected using swallow-tailed perylene derivative fluorescence observed by confocal laser scanning microscopy and high-performance liquid chromatography, respectively. The results obtained from this study warrant further corroboration and study of specific proteins involved in the macrophage activation and their role in inflammations.
MeSH term(s)	Reactive Oxygen Species/metabolism ; Free Radicals/analysis ; Free Radicals/metabolism ; Spin Trapping/methods ; Electron Spin Resonance Spectroscopy/methods ; Macrophages/metabolism ; Proteins/chemistry
Chemical Substances	Reactive Oxygen Species ; Free Radicals ; Proteins
Language	English
Publishing date	2023-02-10
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	60-7
ISSN	1872-8006 ; 1879-2596 ; 1879-260X ; 1879-2642 ; 1879-2618 ; 1879-2650 ; 0006-3002 ; 0005-2728 ; 0005-2736 ; 0304-4165 ; 0167-4838 ; 1388-1981 ; 0167-4889 ; 0167-4781 ; 0304-419X ; 1570-9639 ; 0925-4439 ; 1874-9399
ISSN (online)	1872-8006 ; 1879-2596 ; 1879-260X ; 1879-2642 ; 1879-2618 ; 1879-2650
ISSN	0006-3002 ; 0005-2728 ; 0005-2736 ; 0304-4165 ; 0167-4838 ; 1388-1981 ; 0167-4889 ; 0167-4781 ; 0304-419X ; 1570-9639 ; 0925-4439 ; 1874-9399
DOI	10.1016/j.bbagen.2023.130324
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Article ; Online: Detection and characterization of free radicals induced protein adducts formation in differentiating macrophages

Manoharan, Renuka Ramalingam / Sedlářová, Michaela / Pospisil, Pavel / Prasad, Ankush

BBA - General Subjects. 2023, p.130324-

2023 , Page(s) 130324–

Abstract	Reactive oxygen species play a key role in cellular homeostasis and redox signaling at physiological levels, where excessive production affects the function and integrity of macromolecules, specifically proteins. Therefore, it is important to define free radical-mediated proteotoxic stress in macrophages and identify target protein to prevent tissue dysfunction. A well employed, THP-1 cell line was utilized as in vitro model to study immune response and herein we employ immuno-spin trapping technique to investigate free radical-mediated protein oxidation in macrophages. Hydroxyl radical formation along macrophage differentiation was confirmed by electron paramagnetic resonance along with confocal laser scanning microscopy using hydroxyphenyl fluorescein. Lipid peroxidation product, malondialdehyde, generated under experimental conditions as detected using swallow-tailed perylene derivative fluorescence observed by confocal laser scanning microscopy and high-performance liquid chromatography, respectively. The results obtained from this study warrant further corroboration and study of specific proteins involved in the macrophage activation and their role in inflammations.
Keywords	cell lines ; electron paramagnetic resonance spectroscopy ; fluorescein ; fluorescence ; high performance liquid chromatography ; homeostasis ; hydroxyl radicals ; lipid peroxidation ; macrophage activation ; macrophages ; malondialdehyde ; microscopy ; models ; oxidation ; Reactive oxygen species ; Macrophage ; Phorbol 12-myristate 13-acetate ; All-trans retinoic acid ; Lipopolysaccharide ; Protein oxidation ; ROS ; EPR spectroscopy ; CLSM ; PMA ; RIPA ; ATRA ; LPS ; DNPH ; DIC
Language	English
Publishing place	Elsevier B.V.
Document type	Article ; Online
Note	Pre-press version
ZDB-ID	840755-1
ISSN	0304-4165
ISSN	0304-4165
DOI	10.1016/j.bbagen.2023.130324
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Article ; Online: Differential effects of ascorbic acid on monocytic cell morphology and protein modification: Shifting from pro-oxidative to antioxidant properties.

Prasad, Ankush / Rathi, Deepak / Sedlářová, Michaela / Manoharan, Renuka Ramalingam / Průdková, Eliška / Pospíšil, Pavel

Biochemistry and biophysics reports

2023 Volume 37, Page(s) 101622

Abstract: In this study, we investigated the properties of ascorbic acid (vitamin C), which is a naturally occurring water-soluble vitamin. Our goal is to evaluate its pro-oxidative and/or antioxidant capabilities. To do this, we initially used a confocal laser ... ...

Abstract	In this study, we investigated the properties of ascorbic acid (vitamin C), which is a naturally occurring water-soluble vitamin. Our goal is to evaluate its pro-oxidative and/or antioxidant capabilities. To do this, we initially used a confocal laser scanning microscope (CLSM) to visualize the differentiation pattern in U-937 cells under the treatment of variable concentrations of ascorbic acid. Prior to induction, U-937 cells showed a spherical morphology. After treatment, significant morphological changes were observed in the form of prominent pseudopodia and amoeboid structures. Interestingly, pseudopodia incidences increased with an increase in ascorbic acid concentrations. In addition, our analysis of protein modification using anti-malondialdehyde antibodies showed changes in more than one protein. The findings reveal the link between the differentiation of U-937 cells into macrophages and the protein modifications triggered by the production of reactive oxygen species when U-937 cells are exposed to ascorbic acid. Furthermore, the transformation of ascorbic acid from a pro-oxidative to an antioxidant property is also demonstrated.
Language	English
Publishing date	2023-12-27
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	2831046-9
ISSN	2405-5808 ; 2405-5808
ISSN (online)	2405-5808
ISSN	2405-5808
DOI	10.1016/j.bbrep.2023.101622
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Article ; Online: Formation of free acetaldehydes derived from lipid peroxidation in U937 monocyte-like cells.

Pospíšil, Pavel / Prasad, Ankush / Belková, Julie / Manoharan, Renuka Ramalingam / Sedlářová, Michaela

Biochimica et biophysica acta. General subjects

2023 Volume 1868, Issue 2, Page(s) 130527

Abstract: Acetaldehyde can be found in human cells as a byproduct of various metabolic pathways, including oxidative processes such as lipid peroxidation. This secondary product of lipid peroxidation plays a role in various pathological processes, leading to ... ...

Abstract	Acetaldehyde can be found in human cells as a byproduct of various metabolic pathways, including oxidative processes such as lipid peroxidation. This secondary product of lipid peroxidation plays a role in various pathological processes, leading to various types of civilization diseases. In this study, the formation of free acetaldehyde induced by oxygen-centred radicals was studied in monocyte-like cell line U937. Exposure of U937 cells to peroxyl/alkoxyl radicals induced by azocompound resulted in the formation of free acetaldehyde. Acetaldehyde is formed by the cleavage of fatty acids, which represents the breakdown of fatty acids into smaller fragments initiated by the cyclization of lipid peroxyl radical and β-scission of lipid alkoxyl radical. The cleavage of fatty acids alters the integrity of the plasma and nuclear membrane, leading to the loss of cell viability. Understanding the pathological processes of acetaldehyde formation is an active area of research with potential implications for preventing and treating various diseases associated with oxidative stress.
MeSH term(s)	Humans ; Lipid Peroxidation ; Free Radicals/metabolism ; U937 Cells ; Acetaldehyde ; Monocytes/metabolism ; Fatty Acids/metabolism ; Reactive Oxygen Species
Chemical Substances	Free Radicals ; Acetaldehyde (GO1N1ZPR3B) ; perhydroxyl radical (3170-83-0) ; Fatty Acids ; Reactive Oxygen Species
Language	English
Publishing date	2023-12-02
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	60-7
ISSN	1872-8006 ; 1879-2596 ; 1879-260X ; 1879-2642 ; 1879-2618 ; 1879-2650 ; 0006-3002 ; 0005-2728 ; 0005-2736 ; 0304-4165 ; 0167-4838 ; 1388-1981 ; 0167-4889 ; 0167-4781 ; 0304-419X ; 1570-9639 ; 0925-4439 ; 1874-9399
ISSN (online)	1872-8006 ; 1879-2596 ; 1879-260X ; 1879-2642 ; 1879-2618 ; 1879-2650
ISSN	0006-3002 ; 0005-2728 ; 0005-2736 ; 0304-4165 ; 0167-4838 ; 1388-1981 ; 0167-4889 ; 0167-4781 ; 0304-419X ; 1570-9639 ; 0925-4439 ; 1874-9399
DOI	10.1016/j.bbagen.2023.130527
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Article ; Online: Free Radical-Mediated Protein Radical Formation in Differentiating Monocytes.

Prasad, Ankush / Manoharan, Renuka Ramalingam / Sedlářová, Michaela / Pospíšil, Pavel

International journal of molecular sciences

2021 Volume 22, Issue 18

Abstract: Free radical-mediated activation of inflammatory macrophages remains ambiguous with its limitation to study within biological systems. U-937 and HL-60 cell lines serve as a well-defined model system known to differentiate into either macrophages or ... ...

Abstract	Free radical-mediated activation of inflammatory macrophages remains ambiguous with its limitation to study within biological systems. U-937 and HL-60 cell lines serve as a well-defined model system known to differentiate into either macrophages or dendritic cells in response to various chemical stimuli linked with reactive oxygen species (ROS) production. Our present work utilizes phorbol 12-myristate-13-acetate (PMA) as a stimulant, and factors such as concentration and incubation time were considered to achieve optimized differentiation conditions. ROS formation likely hydroxyl radical (HO
MeSH term(s)	Acetophenones/pharmacology ; Cell Differentiation/drug effects ; Cell Proliferation/drug effects ; Cell Shape/drug effects ; Cell Survival/drug effects ; Electron Spin Resonance Spectroscopy ; Free Radicals/metabolism ; HL-60 Cells ; Humans ; Hydroxyl Radical ; Monocytes/cytology ; Monocytes/drug effects ; Monocytes/metabolism ; NADP/metabolism ; Proteins/metabolism ; Staining and Labeling ; Tetradecanoylphorbol Acetate/pharmacology ; U937 Cells
Chemical Substances	Acetophenones ; Free Radicals ; Proteins ; Hydroxyl Radical (3352-57-6) ; NADP (53-59-8) ; acetovanillone (B6J7B9UDTR) ; Tetradecanoylphorbol Acetate (NI40JAQ945)
Language	English
Publishing date	2021-09-15
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms22189963
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Article: Re-Evaluation of Imaging Methods of Reactive Oxygen and Nitrogen Species in Plants and Fungi: Influence of Cell Wall Composition.

Sedlářová, Michaela / Luhová, Lenka

Frontiers in physiology

2017 Volume 8, Page(s) 826

Abstract: Developmental transitions and stress reactions in both eukaryotes and prokaryotes are tightly linked with fast and localized modifications in concentrations of reactive oxygen and nitrogen species (ROS and RNS). Fluorescent microscopic analyses are ... ...

Abstract	Developmental transitions and stress reactions in both eukaryotes and prokaryotes are tightly linked with fast and localized modifications in concentrations of reactive oxygen and nitrogen species (ROS and RNS). Fluorescent microscopic analyses are widely applied to detect localized production of ROS and RNS
Language	English
Publishing date	2017-10-24
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2564217-0
ISSN	1664-042X
ISSN	1664-042X
DOI	10.3389/fphys.2017.00826
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Article ; Online: Genetic structure of Plasmopara halstedii populations across Europe and South Russia

Kitner, Miloslav / Thines, Marco / Sedlářová, Michaela / Vaculná, Lucie / Bán, Rita / Körösi, Katalin / Iwebor, Maria / Antonova, Tatiana / Ali, Tahir / Nádvorník, Petr / Lebeda, A. / Spring, Otmar

Plant Pathology. 2023 Feb., v. 72, no. 2 p.361-375

2023

Abstract: Plasmopara halstedii is a biotrophic parasite of sunflower (Helianthus annuus). We genetically analysed 162 P. halstedii samples collected between 1982 and 2018, representing a 3500 km west–east transect across Europe, from France to the Krasnodar region ...

Abstract	Plasmopara halstedii is a biotrophic parasite of sunflower (Helianthus annuus). We genetically analysed 162 P. halstedii samples collected between 1982 and 2018, representing a 3500 km west–east transect across Europe, from France to the Krasnodar region in southern Russia. To assess the population genetic structure among P. halstedii isolates, we sequenced two mitochondrial regions (cox1 and cox2), in addition to analysing eight polymorphic simple sequence repeat (SSR, microsatellite) markers. Sequencing of cox2 enabled comparison of our data with recent SSR‐based studies to provide further evidence for differences among P. halstedii strains infecting different hosts (H. annuus, H. × laetiflorus, Rudbeckia spp.). There are two different lineages infecting H. annuus that we resolved on a neighbour‐joining tree, which also corresponded to a discriminant analysis of principal components. This suggests at least two independent introductions of P. halstedii on sunflower. We observed no relationship between the phylogenetic placement of the individual samples and virulence characteristics, indicating independent evolution of virulence phenotypes. Both genetic markers clearly separated P. halstedii samples originating from H. annuus and H. × laetiflorus, suggesting these might represent distinct species.
Keywords	Helianthus annuus ; Plasmopara halstedii ; Rudbeckia ; Russia ; discriminant analysis ; genetic structure ; microsatellite repeats ; mitochondria ; parasites ; phylogeny ; plant pathology ; population genetics ; virulence ; France
Language	English
Dates of publication	2023-02
Size	p. 361-375.
Publishing place	John Wiley & Sons, Ltd
Document type	Article ; Online
Note	JOURNAL ARTICLE
ZDB-ID	415941-x
ISSN	1365-3059 ; 0032-0862
ISSN (online)	1365-3059
ISSN	0032-0862
DOI	10.1111/ppa.13666
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Article: Characterization of Protein Radicals in Arabidopsis.

Kumar, Aditya / Prasad, Ankush / Sedlářová, Michaela / Pospíšil, Pavel

Frontiers in physiology

2019 Volume 10, Page(s) 958

Abstract: Oxidative modification of proteins in photosystem II (PSII) exposed to high light has been studied for a few decades, but the characterization of protein radicals formed by protein oxidation is largely unknown. Protein oxidation is induced by the direct ... ...

Abstract	Oxidative modification of proteins in photosystem II (PSII) exposed to high light has been studied for a few decades, but the characterization of protein radicals formed by protein oxidation is largely unknown. Protein oxidation is induced by the direct reaction of proteins with reactive oxygen species known to form highly reactive protein radicals comprising carbon-centered (alkyl) and oxygen-centered (peroxyl and alkoxyl) radicals. In this study, protein radicals were monitored in Arabidopsis exposed to high light by immuno-spin trapping technique based on the detection of 5,5-dimethyl-1-pyrroline N-oxide (DMPO) nitrone adducts using the anti-DMPO antibody. Protein radicals were imaged in Arabidopsis leaves and chloroplasts by confocal laser scanning microscopy using fluorescein conjugated with the anti-DMPO antibody. Characterization of protein radicals by standard blotting techniques using PSII protein specific antibodies shows that protein radicals are formed on D1, D2, CP43, CP47, and Lhcb3 proteins. Protein oxidation reflected by the appearance/disappearance of the protein bands reveals that formation of protein radicals was associated with protein fragmentation (cleavage of the D1 peptide bonds) and aggregation (cross-linking with another PSII subunits). Characterization of protein radical formation is important for better understating of the mechanism of oxidative modification of PSII proteins under high light.
Language	English
Publishing date	2019-08-13
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2564217-0
ISSN	1664-042X
ISSN	1664-042X
DOI	10.3389/fphys.2019.00958
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