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Article ; Online: ROS signaling in innate immunity via oxidative protein modifications.

Manoharan, Renuka Ramalingam / Prasad, Ankush / Pospíšil, Pavel / Kzhyshkowska, Julia

2024 Volume 15, Page(s) 1359600

Abstract: The innate immune response represents the first-line of defense against invading pathogens. Reactive oxygen species (ROS) and reactive nitrogen species (RNS) have been implicated in various aspects of innate immune function, which involves respiratory ... ...

Abstract	The innate immune response represents the first-line of defense against invading pathogens. Reactive oxygen species (ROS) and reactive nitrogen species (RNS) have been implicated in various aspects of innate immune function, which involves respiratory bursts and inflammasome activation. These reactive species widely distributed within the cellular environment are short-lived intermediates that play a vital role in cellular signaling and proliferation and are likely to depend on their subcellular site of formation. NADPH oxidase complex of phagocytes is known to generate superoxide anion radical (O
MeSH term(s)	Hydrogen Peroxide ; Oxidation-Reduction ; Superoxides ; Oxidative Stress ; Hypochlorous Acid ; Immunity, Innate
Chemical Substances	Hydrogen Peroxide (BBX060AN9V) ; Superoxides (11062-77-4) ; Hypochlorous Acid (712K4CDC10)
Language	English
Publishing date	2024-03-07
Publishing country	Switzerland
Document type	Journal Article ; Review ; Research Support, Non-U.S. Gov't
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2024.1359600
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Article ; Online: Malondialdehyde enhances PsbP protein release during heat stress in Arabidopsis.

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

Plant physiology and biochemistry : PPB

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: Reactive oxygen species in photosystem II: relevance for oxidative signaling

Pospisil, Pavel / Kumar, Aditya / Prasad, Ankush

Photosynth Res. 2022 June, v. 152, no. 3 p.245-260

2022

Abstract: Reactive oxygen species (ROS) are formed in photosystem II (PSII) under various types of abiotic and biotic stresses. It is considered that ROS play a role in chloroplast-to-nucleus retrograde signaling, which changes the nuclear gene expression. However, ...

Abstract	Reactive oxygen species (ROS) are formed in photosystem II (PSII) under various types of abiotic and biotic stresses. It is considered that ROS play a role in chloroplast-to-nucleus retrograde signaling, which changes the nuclear gene expression. However, as ROS lifetime and diffusion are restricted due to the high reactivity towards biomolecules (lipids, pigments, and proteins) and the spatial specificity of signal transduction is low, it is not entirely clear how ROS might transduce signal from the chloroplasts to the nucleus. Biomolecule oxidation was formerly connected solely with damage; nevertheless, the evidence appears that oxidatively modified lipids and pigments are be involved in chloroplast-to-nucleus retrograde signaling due to their long diffusion distance. Moreover, oxidatively modified proteins show high spatial specificity; however, their role in signal transduction from chloroplasts to the nucleus has not been proven yet. The review attempts to summarize and evaluate the evidence for the involvement of ROS in oxidative signaling in PSII.
Keywords	biochemical compounds ; chloroplasts ; gene expression ; oxidation ; photosystem II ; reactive oxygen species ; signal transduction
Language	English
Dates of publication	2022-06
Size	p. 245-260.
Publishing place	Springer Netherlands
Document type	Article ; Online
Note	Review
ZDB-ID	1475688-2
ISSN	1573-5079 ; 0166-8595
ISSN (online)	1573-5079
ISSN	0166-8595
DOI	10.1007/s11120-022-00922-x
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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: Scanning Electrochemical Microscopy-Somatic Cell Count as a Method for Diagnosis of Bovine Mastitis.

Kasai, Shigenobu / Prasad, Ankush / Kumagai, Ryoma / Takanohashi, Keita

Biology

2022 Volume 11, Issue 4

Abstract: The method to diagnose mastitis is generally the somatic cell count (SCC) by flow cytometry measurement. When the number of somatic cells in raw milk is 2.0 × ... ...

Abstract	The method to diagnose mastitis is generally the somatic cell count (SCC) by flow cytometry measurement. When the number of somatic cells in raw milk is 2.0 × 10
Language	English
Publishing date	2022-04-01
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2661517-4
ISSN	2079-7737
ISSN	2079-7737
DOI	10.3390/biology11040549
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Article ; Online: Reactive oxygen species in photosystem II: relevance for oxidative signaling.

Pospíšil, Pavel / Kumar, Aditya / Prasad, Ankush

Photosynthesis research

2022 Volume 152, Issue 3, Page(s) 245–260

Abstract	Reactive oxygen species (ROS) are formed in photosystem II (PSII) under various types of abiotic and biotic stresses. It is considered that ROS play a role in chloroplast-to-nucleus retrograde signaling, which changes the nuclear gene expression. However, as ROS lifetime and diffusion are restricted due to the high reactivity towards biomolecules (lipids, pigments, and proteins) and the spatial specificity of signal transduction is low, it is not entirely clear how ROS might transduce signal from the chloroplasts to the nucleus. Biomolecule oxidation was formerly connected solely with damage; nevertheless, the evidence appears that oxidatively modified lipids and pigments are be involved in chloroplast-to-nucleus retrograde signaling due to their long diffusion distance. Moreover, oxidatively modified proteins show high spatial specificity; however, their role in signal transduction from chloroplasts to the nucleus has not been proven yet. The review attempts to summarize and evaluate the evidence for the involvement of ROS in oxidative signaling in PSII.
MeSH term(s)	Chloroplasts/metabolism ; Lipids ; Oxidative Stress ; Photosystem II Protein Complex/metabolism ; Reactive Oxygen Species/metabolism ; Signal Transduction/physiology
Chemical Substances	Lipids ; Photosystem II Protein Complex ; Reactive Oxygen Species
Language	English
Publishing date	2022-05-28
Publishing country	Netherlands
Document type	Journal Article ; Review
ZDB-ID	1475688-2
ISSN	1573-5079 ; 0166-8595
ISSN (online)	1573-5079
ISSN	0166-8595
DOI	10.1007/s11120-022-00922-x
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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
Database	NAL-Catalogue (AGRICOLA)

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Article: NOX2 and NOX4 expression in monocytes and macrophages-extracellular vesicles in signalling and therapeutics.

Rathi, Deepak / Rossi, Claudio / Pospíšil, Pavel / Ramalingam Manoharan, Renuka / Talarico, Luigi / Magnani, Agnese / Prasad, Ankush

Frontiers in cell and developmental biology

2024 Volume 12, Page(s) 1342227

Abstract: Extracellular vesicles (EVs) are a type of cytoplasmic vesicles secreted by a variety of cells. EVs originating from cells have been known to participate in cell communication, antigen presentation, immune cell activation, tolerance induction, etc. These ...

Abstract	Extracellular vesicles (EVs) are a type of cytoplasmic vesicles secreted by a variety of cells. EVs originating from cells have been known to participate in cell communication, antigen presentation, immune cell activation, tolerance induction, etc. These EVs can also carry the active form of Nicotinamide Adenine Dinucleotide Phosphate Oxidase Hydrogen (NADPH) oxidase, which is very essential for the production of reactive oxygen species (ROS) and that can then modulate processes such as cell regeneration. The aim of this study is to characterize the EVs isolated from U-937 and THP-1 cells, identify the NADPH oxidase (NOX) isoforms, and to determine whether EVs can modulate NOX4 and NOX2 in monocytes and macrophages. In our study, isolated EVs of U-937 were characterized using dynamic light scattering (DLS) spectroscopy and immunoblotting. The results showed that the exogenous addition of differentiation agents (either phorbol 12-myristate 13-acetate (PMA) or ascorbic acid) or the supplementation of EVs used in the study did not cause any stress leading to alterations in cell proliferation and viability. In cells co-cultured with EVs for 72 h, strong suppression of NOX4 and NOX2 is evident when monocytes transform into macrophagic cells. We also observed lower levels of oxidative stress measured using immunoblotting and electron paramagnetic resonance spectroscopy under the EVs co-cultured condition, which also indicates that EVs might contribute significantly by acting as an antioxidant source, which agrees with previous studies that hypothesized the role of EVs in therapeutics. Therefore, our results provide evidence for NOX regulation by EVs in addition to its role as an antioxidant cargo.
Language	English
Publishing date	2024-04-16
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2737824-X
ISSN	2296-634X
ISSN	2296-634X
DOI	10.3389/fcell.2024.1342227
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Article ; Online: Imaging and Characterization of Oxidative Protein Modifications in Skin.

Prasad, Ankush / Duchová, Hana / Manoharan, Renuka Ramalingam / Rathi, Deepak / Pospíšil, Pavel

International journal of molecular sciences

2023 Volume 24, Issue 4

Abstract: Skin plays an important role in protection, metabolism, thermoregulation, sensation, and excretion whilst being consistently exposed to environmental aggression, including biotic and abiotic stresses. During the generation of oxidative stress in the skin, ...

Abstract	Skin plays an important role in protection, metabolism, thermoregulation, sensation, and excretion whilst being consistently exposed to environmental aggression, including biotic and abiotic stresses. During the generation of oxidative stress in the skin, the epidermal and dermal cells are generally regarded as the most affected regions. The participation of reactive oxygen species (ROS) as a result of environmental fluctuations has been experimentally proven by several researchers and is well known to contribute to ultra-weak photon emission via the oxidation of biomolecules (lipids, proteins, and nucleic acids). More recently, ultra-weak photon emission detection techniques have been introduced to investigate the conditions of oxidative stress in various living systems in in vivo, ex vivo and in vitro studies. Research into two-dimensional photon imaging is drawing growing attention because of its application as a non-invasive tool. We monitored spontaneous and stress-induced ultra-weak photon emission under the exogenous application of a Fenton reagent. The results showed a marked difference in the ultra-weak photon emission. Overall, these results suggest that triplet carbonyl (
MeSH term(s)	Reactive Oxygen Species/metabolism ; Hydrogen Peroxide/metabolism ; Skin/metabolism ; Oxidative Stress ; Oxidation-Reduction ; Proteins/metabolism
Chemical Substances	Reactive Oxygen Species ; Hydrogen Peroxide (BBX060AN9V) ; Proteins
Language	English
Publishing date	2023-02-16
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/ijms24043981
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