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Acta Biochimica Et Biophysica Sinica Jul 2022In the present study, we investigate the effect of homocysteine (Hcy) on extracellular-superoxide dismutase (EC-SOD) DNA methylation in the aorta of mice, and explore...
In the present study, we investigate the effect of homocysteine (Hcy) on extracellular-superoxide dismutase (EC-SOD) DNA methylation in the aorta of mice, and explore the underlying mechanism in macrophages, trying to identify the key targets of Hcy-induced EC-SOD methylation changes. mice are fed different diets for 15 weeks, EC-SOD and DNA methyltransferase 1 (DNMT1) expression levels are detected by RT-PCR and western blot analysis. EC-SOD methylation levels are assessed by ntMS-PCR. After EC-SOD overexpression or knockdown in macrophages, following the transfection of macrophages with pEGFP-N1-DNMT1, the methylation levels of EC-SOD are detected. Our data show that the concentrations of Hcy and the area of atherogenic lesions are significantly increased in mice fed with a high-methionine diet, and have a positive correlation with the levels of superoxide anions, which indicates that Hcy-activated superoxide anions enhance the development of atherogenic lesions. EC-SOD expression is suppressed by Hcy, and the content of superoxide anion is increased when EC-SOD is silenced by RNAi in macrophages, suggesting that EC-SOD plays a major part in oxidative stress induced by Hcy. Furthermore, the promoter activity of EC-SOD is increased following transfection with the -1/-1100 fragment, and EC-SOD methylation level is significantly suppressed by Hcy, and more significantly decreased upon DNMT1 overexpression. In conclusion, Hcy may alter the DNA methylation status and DNMT1 acts as the essential enzyme in the methyl transfer process to disturb the status of EC-SOD DNA methylation, leading to decreased expression of EC-SOD and increased oxidative stress and atherosclerosis.
Topics: Mice; Animals; DNA Methylation; Superoxides; Homocysteine; Atherosclerosis; Superoxide Dismutase; Oxidative Stress; Apolipoproteins E
PubMed: 35866603
DOI: 10.3724/abbs.2022093 -
International Journal of Molecular... Feb 2023Copper-zinc superoxide dismutase 1 (SOD1) has long been recognized as a major redox enzyme in scavenging superoxide radicals. However, there is little information on its...
Copper-zinc superoxide dismutase 1 (SOD1) has long been recognized as a major redox enzyme in scavenging superoxide radicals. However, there is little information on its non-canonical role and metabolic implications. Using a protein complementation assay (PCA) and pull-down assay, we revealed novel protein-protein interactions (PPIs) between SOD1 and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (YWHAZ) or epsilon (YWHAE) in this research. Through site-directed mutagenesis of SOD1, we studied the binding conditions of the two PPIs. Forming the SOD1 and YWHAE or YWHAZ protein complex enhanced enzyme activity of purified SOD1 in vitro by 40% ( < 0.05) and protein stability of over-expressed intracellular YWHAE (18%, < 0.01) and YWHAZ (14%, < 0.05). Functionally, these PPIs were associated with lipolysis, cell growth, and cell survival in HEK293T or HepG2 cells. In conclusion, our findings reveal two new PPIs between SOD1 and YWHAE or YWHAZ and their structural dependences, responses to redox status, mutual impacts on the enzyme function and protein degradation, and metabolic implications. Overall, our finding revealed a new unorthodox role of SOD1 and will provide novel perspectives and insights for diagnosing and treating diseases related to the protein.
Topics: Humans; Copper; HEK293 Cells; Superoxide Dismutase; Superoxide Dismutase-1; Superoxides
PubMed: 36834640
DOI: 10.3390/ijms24043230 -
F1000Research 2023Superoxide dismutase [Cu-Zn] 1 (SOD1), is an antioxidant enzyme encoded by the gene , responsible for regulating oxidative stress levels by sequestering free radicals....
Superoxide dismutase [Cu-Zn] 1 (SOD1), is an antioxidant enzyme encoded by the gene , responsible for regulating oxidative stress levels by sequestering free radicals. Identified as the first gene with mutations in Amyotrophic lateral sclerosis (ALS), is a determinant for studying diseases of aging and neurodegeneration. With guidance on well-characterized anti-SOD1 antibodies, the reproducibility of SOD1 research would be enhanced. In this study, we characterized eleven SOD1 commercial antibodies for Western blot, immunoprecipitation, and immunofluorescence using a standardized experimental protocol based on comparing read-outs in knockout cell lines and isogenic parental controls. We identified many high-performing antibodies and encourage readers to use this report as a guide to select the most appropriate antibody for their specific needs.
Topics: Superoxide Dismutase-1; Reproducibility of Results; Superoxide Dismutase; Blotting, Western; Antibodies; Immunoprecipitation; Fluorescent Antibody Technique; Zinc
PubMed: 37860271
DOI: 10.12688/f1000research.132952.1 -
Arhiv Za Higijenu Rada I Toksikologiju Sep 2020Oxidative stress occurs when reactive oxygen species (ROS) production overwhelms cell protection by antioxidants. This review is focused on general anaesthesia-induced... (Review)
Review
Oxidative stress occurs when reactive oxygen species (ROS) production overwhelms cell protection by antioxidants. This review is focused on general anaesthesia-induced oxidative stress because it increases the rate of complications and delays recovery after surgery. It is important to know what effects of anaesthetics to expect in terms of oxidative stress, particularly in surgical procedures with high ROS production, because their either additive or antagonistic effect may be pivotal for the outcome of surgery. In vitro and animal studies on this topic are numerous but show large variability. There are not many human studies and what we know has been learned from different surgical procedures measuring different endpoints in blood samples taken mostly before and after surgery. In these studies most intravenous anaesthetics have antioxidative properties, while volatile anaesthetics temporarily increase oxidative stress in longer surgical procedures.
Topics: Anesthesia, Inhalation; Animals; Antioxidants; Humans; Oxidative Stress; Reactive Oxygen Species; Superoxide Dismutase
PubMed: 33074169
DOI: 10.2478/aiht-2020-71-3437 -
International Journal of Molecular... Jun 2021Quantum dots (QDs) have a broad range of applications in cell biolabeling, cancer treatment, metastasis imaging, and therapeutic drug monitoring. Despite their wide use,...
Quantum dots (QDs) have a broad range of applications in cell biolabeling, cancer treatment, metastasis imaging, and therapeutic drug monitoring. Despite their wide use, relatively little is known about their influence on other molecules. Interactions between QDs and proteins can influence the properties of both nanoparticles and proteins. The effect of mercaptosuccinic acid-capped CdTe QDs on intercellular copper-zinc superoxide dismutase (SOD1)-one of the main enzymatic antioxidants-was investigated. Incubation of SOD1 with QDs caused an increase in SOD1 activity, unlike in the case of CdCl, which inhibited SOD1. Moreover, this effect on SOD1 increased with the size and potential of QDs, although the effect became clearly visible in higher concentrations of QDs. The intensity of QD-SOD1 fluorescence, analyzed with the use of capillary electrophoresis with laser-induced fluorescence detection, was dependent on SOD1 concentration. In the case of green QDs, the fluorescence signal decreased with increasing SOD1 concentration. In contrast, the signal strength for Y-QD complexes was not dependent on SOD1 dilutions. The migration time of QDs and their complexes with SOD1 varied depending on the type of QD used. The migration time of G-QD complexes with SOD1 differed slightly. However, in the case of Y-QD complexes with SOD1, the differences in the migration time were not dependent on SOD concentration. This research shows that QDs interact with SOD1 and the influence of QDs on SOD activity is size-dependent. With this knowledge, one might be able to control the activation/inhibition of specific enzymes, such as SOD1.
Topics: Cadmium Compounds; Electrophoresis, Capillary; Fluorescence; Humans; Nanoparticles; Quantum Dots; Spectrometry, Fluorescence; Superoxide Dismutase; Tellurium
PubMed: 34200401
DOI: 10.3390/ijms22116156 -
Bulletin of Experimental Biology and... Jan 2022The study examined the skin histomorphology and biochemistry in mature ovariectomized rats treated and not treated with estrogen. Biochemical parameters (superoxide...
The study examined the skin histomorphology and biochemistry in mature ovariectomized rats treated and not treated with estrogen. Biochemical parameters (superoxide dismutase, malondialdehyde, and hydroxyproline content) were measured in dorsal skin samples collected in 50 days after surgery. The morphology of dorsal skin was analyzed under a microscope. In ovariectomized rats, the skin levels of superoxide dismutase and hydroxyproline were significantly lower, while the superoxide dismutase content was significantly higher than in shamoperated animals (p<0.05). Estrogen therapy significantly increased the levels of superoxide dismutase and hydroxyproline and reduced superoxide dismutase level in ovariectomized rats in comparison with the corresponding parameters in untreated ovariectomized animals (p<0.05). Histomorphological analysis of the skin from non-treated ovariectomized rats revealed reduced vascularization and lower density of papillary capillaries in comparison with these parameters in sham-operated controls; estrogen treatment prevented these changes. We concluded that ovariectomized rats can be employed as a model of aging skin in menopause.
Topics: Animals; Female; Humans; Malondialdehyde; Menopause; Ovariectomy; Oxidative Stress; Rats; Skin Aging; Superoxide Dismutase
PubMed: 35001317
DOI: 10.1007/s10517-022-05396-4 -
Animal Biotechnology Dec 2023The aim of this study is to determine the effects of 50% of 96 h LC (5.25 ppm) diazinon on the expression of superoxide dismutase (SOD) enzyme genes (, , and ) and...
The aim of this study is to determine the effects of 50% of 96 h LC (5.25 ppm) diazinon on the expression of superoxide dismutase (SOD) enzyme genes (, , and ) and SOD enzyme activity at the end of 24, 48, 72, and 96 h in platyfish liver and gill tissues. To this end, we determined the tissue-specific distribution of , , and genes and performed analyses in platyfish (). It was determined that malondialdehyde (MDA) level and SOD enzyme activity were increased in the liver [(43.90 EU mg protein (control), 62.45 EU mg protein (24 h), 73.17 EU mg protein (48 h), 82.18 EU mg protein (72 h), 92.93 EU mg protein (96 h)] and gill [(16.44 EU mg protein (control), 33.47 EU mg protein (24 h), 50.38 EU mg protein (48 h), 64.62 EU mg protein (72 h), 74.04 EU mg protein (96 h)] tissues of platyfish exposed to diazinon, while the expression of the genes was down-regulated. The tissue-specific distribution of the genes varied, with the tissues and the genes expression were being predominant in the liver (628.32 in , 637.59 in , 888.5 in ). Thus, the liver was considered a suitable tissue for further gene expression studies. Based on the phylogenetic analyses, platyfish genes can be reported to be orthologs of / genes from other vertebrates. Identity/similarity analyses supported this determination. Conserved gene synteny proved that there are conserved genes in platyfish, zebrafish, and humans.
Topics: Humans; Animals; Diazinon; Phylogeny; Zebrafish; Superoxide Dismutase; Superoxide Dismutase-1; Genomics; Cyprinodontiformes
PubMed: 36811494
DOI: 10.1080/10495398.2023.2178931 -
Redox Biology May 2020Extracellular superoxide dismutase (EcSOD) is the only extracellular scavenger of superoxide anion (O) with unique binding capacity to cell surface and extracellular... (Review)
Review
Extracellular superoxide dismutase (EcSOD) is the only extracellular scavenger of superoxide anion (O) with unique binding capacity to cell surface and extracellular matrix through its heparin-binding domain. Enhanced EcSOD activity prevents oxidative stress and damage, which are fundamental in a variety of disease pathologies. In this review we will discuss the findings in humans and animal studies supporting the benefits of EcSOD induced by exercise training in reducing oxidative stress in various tissues. In particularly, we will highlight the importance of skeletal muscle EcSOD, which is induced by endurance exercise and redistributed through the circulation to the peripheral tissues, as a molecular transducer of exercise training to confer protection against oxidative stress and damage in various disease conditions.
Topics: Animals; Exercise; Humans; Oxidative Stress; Superoxide Dismutase; Superoxides; Transducers
PubMed: 32220789
DOI: 10.1016/j.redox.2020.101508 -
Scientific Reports Apr 2023A novel hybrid protein composed of a superoxide dismutase-active Cu(II) complex (CuST) and lysozyme (CuST@lysozyme) was prepared. The results of the spectroscopic and...
A novel hybrid protein composed of a superoxide dismutase-active Cu(II) complex (CuST) and lysozyme (CuST@lysozyme) was prepared. The results of the spectroscopic and electrochemical analyses confirmed that CuST binds to lysozyme. We determined the crystal structure of CuST@lysozyme at 0.92 Å resolution, which revealed that the His15 imidazole group of lysozyme binds to the Cu(II) center of CuST in the equatorial position. In addition, CuST was fixed in position by the weak axial coordination of the Thr89 hydroxyl group and the hydrogen bond between the guanidinium group of the Arg14 residue and the hydroxyl group of CuST. Furthermore, the combination of CuST with lysozyme did not decrease the superoxide dismutase activity of CuST. Based on the spectral, electrochemical, structural studies, and quantum chemical calculations, an O disproportionation mechanism catalyzed by CuST@lysozyme is proposed.
Topics: Superoxide Dismutase; Superoxides; Oxidation-Reduction; Muramidase; Copper
PubMed: 37106030
DOI: 10.1038/s41598-023-33926-1 -
Molecular Biology Reports Nov 2020Superoxide dismutase (SOD) is an antioxidant enzyme that acts as a component of first-line defense system against reactive oxygen species (ROS). Copper/Zinc superoxide...
Superoxide dismutase (SOD) is an antioxidant enzyme that acts as a component of first-line defense system against reactive oxygen species (ROS). Copper/Zinc superoxide dismutase (Cu/Zn-SOD) is one of the isoforms of SOD enzyme and is sensitive to the exposure of different environmental factors, in different species and tissues. Caiman latirostris is one of the two crocodilian species living in Argentina and no information is available on the molecular and biochemical characteristics of the Cu/Zn-sod gene in this species. In the present work, we reported the presence of the Cu/Zn-sod gene in C. latirostris, the nucleotide and amino acid sequences, the modelled protein structure, evolutionary distance among species and tissue specific expression patterns. Cu/Zn-sod gene was 620 bp open reading frame in length and encoded 178 amino acids. The nucleotide sequences of C. latirostris shared high similarity with the Cu/Zn-sod genes of other crocodilian species, so it showed to be highly conserved. PCR analysis showed that Cu/Zn-sod gene was expressed in all the tissues examined (liver, gonads, spleen, heart, and whole blood), suggesting a constitutively expressed gene in these tissues. This study allows further investigation into the structure-activity relationship and the mechanism of action of Cu/Zn-SOD, besides exploring the functional breadth and possible alteration factors, including xenobiotics.
Topics: Alligators and Crocodiles; Amino Acid Sequence; Animals; Argentina; Computational Biology; Gene Expression Profiling; Gene Expression Regulation, Enzymologic; Models, Molecular; Phylogeny; Protein Structure, Tertiary; Sequence Homology, Amino Acid; Superoxide Dismutase-1
PubMed: 33128685
DOI: 10.1007/s11033-020-05937-y