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Protein and Peptide Letters 2023Designing effective diagnostics, biotherapeutics, and biocatalysts are a few interesting potential outcomes of protein engineering. Despite being just a few decades old,... (Review)
Review
Designing effective diagnostics, biotherapeutics, and biocatalysts are a few interesting potential outcomes of protein engineering. Despite being just a few decades old, the discipline of de novo protein designing has provided a foundation for remarkable outcomes in the pharmaceuticals and enzyme industries. The technologies that will have the biggest impact on current protein therapeutics include engineered natural protein variants, Fc fusion protein, and antibody engineering. Furthermore, designing protein scaffolds can be used in developing next-generation antibodies and in transplanting active sites in the enzyme. The article highlights the important tools and techniques used in protein engineering and their application in the engineering of enzymes and therapeutic proteins. This review further sheds light on the engineering of superoxide dismutase, an enzyme responsible for catalyzing the conversion of superoxide radicals to oxygen and hydrogen peroxide by catalyzing a redox reaction at the metal center while concurrently oxidizing and reducing superoxide free radicals.
Topics: Proteins; Protein Engineering; Superoxide Dismutase; Antibodies; Oxidation-Reduction
PubMed: 37211849
DOI: 10.2174/0929866530666230519122612 -
Molecular Metabolism Sep 2022Mutations in the copper-zinc superoxide dismutase (SOD1) gene cause familial amyotrophic lateral sclerosis (ALS), a progressive fatal neuromuscular disease characterized...
OBJECTIVE
Mutations in the copper-zinc superoxide dismutase (SOD1) gene cause familial amyotrophic lateral sclerosis (ALS), a progressive fatal neuromuscular disease characterized by motor neurons death and severe skeletal muscle degeneration. However, there is no effective treatment for this debilitating disease, since the underlying cause for the pathogenesis remains poorly understood. Here, we investigated a role of acyl-CoA:lysocardiolipin acyltransferase 1 (ALCAT1), an acyltransferase that promotes mitochondrial dysfunction in age-related diseases by catalyzing pathological remodeling of cardiolipin, in promoting the development of ALS in the SOD1 transgenic mice.
METHODS
Using SOD1 transgenic mice with targeted deletion of the ALCAT1 gene and treated with Dafaglitapin (Dafa), a very potent and highly selective ALCAT1 inhibitor, we determined whether ablation or pharmaceutical inhibition of ALCAT1 by Dafa would mitigate ALS and the underlying pathogenesis by preventing pathological remodeling of cardiolipin, oxidative stress, and mitochondrial dysfunction by multiple approaches, including lifespan analysis, behavioral tests, morphological and functional analysis of skeletal muscle, electron microscopic and Seahorse analysis of mitochondrial morphology and respiration, western blot analysis of the SOD1 protein aggregation, and lipidomic analysis of cardiolipin content and acyl composition in mice spinal cord.
RESULTS
ALCAT1 protein expression is potently upregulated in the skeletal muscle of the SOD1 mice. Consequently, ablation or pharmacological inhibition of ALCAT1 by Dafa attenuates motor neuron dysfunction, neuronal inflammation, and skeletal muscle atrophy in SOD1 mice by preventing SOD1 protein aggregation, mitochondrial dysfunction, and pathological CL remodeling, leading to moderate extension of lifespan in the SOD1 transgenic mice.
CONCLUSIONS
ALCAT1 promotes the development of ALS by linking SOD1 protein aggregation to mitochondrial dysfunction, implicating Dafa as a potential treatment for this debilitating disorder.
Topics: Acyltransferases; Amyotrophic Lateral Sclerosis; Animals; Cardiolipins; Disease Models, Animal; Mice; Mice, Transgenic; Protein Aggregates; Superoxide Dismutase; Superoxide Dismutase-1
PubMed: 35772643
DOI: 10.1016/j.molmet.2022.101536 -
Journal of Neural Transmission (Vienna,... Jun 2022Autism spectrum disorder is a pervasive neurodevelopmental disorder with a substantial contribution to the global disease burden. Despite intensive research efforts, the...
Autism spectrum disorder is a pervasive neurodevelopmental disorder with a substantial contribution to the global disease burden. Despite intensive research efforts, the aetiopathogenesis remains unclear. The Janus-faced antioxidant enzymes superoxide dismutase 1-3 have been implicated in initiating oxidative stress and as such may constitute a potential therapeutic target. However, no measurement has been taken in human autistic brain samples. The aim of this study is to measure superoxide dismutase 1-3 in autistic cerebral organoids as an in vitro model of human foetal neurodevelopment. Whole brain organoids were created from induced pluripotent stem cells from healthy individuals (n = 5) and individuals suffering from autism (n = 4). Using Pierce bicinchoninic acid and enzyme-linked immunosorbent assays, the protein and superoxide dismutase 1, 2, and 3 concentrations were quantified in the cerebral organoids at days 22, 32, and 42. Measurements were normalized to the protein concentration. Results represented using medians and interquartile ranges. Using Wilcoxon matched-pairs signed-rank test, an abrupt rise in the superoxide dismutase concentration was observed at day 32 and onwards. Using Wilcoxon rank-sum test, no differences were observed between healthy (SOD1: 35.56 ng/mL ± 3.46; SOD2: 2435.80 ng/mL ± 1327.00; SOD3: 1854.88 ng/mL ± 867.94) and autistic (SOD1: 32.85 ng/mL ± 5.26; SOD2: 2717.80 ng/mL ± 1889.10; SOD3: 1690.18 ng/mL ± 615.49) organoids. Cerebral organoids recapitulate many aspects of human neurodevelopment, but the diffusion restriction may render efforts in modelling differences in oxidative stress futile due to the intrinsic hypoxia and central necrosis.
Topics: Autism Spectrum Disorder; Humans; Isoenzymes; Organoids; Oxidative Stress; Superoxide Dismutase; Superoxide Dismutase-1
PubMed: 35266053
DOI: 10.1007/s00702-022-02472-x -
Fish & Shellfish Immunology Jun 2024Copper/zinc superoxide dismutase (Cu/Zn-SOD) can effectively eliminate reactive oxygen species (ROS),avoid damage from O to the body, and maintain O balance. In this...
Copper/zinc superoxide dismutase (Cu/Zn-SOD) can effectively eliminate reactive oxygen species (ROS),avoid damage from O to the body, and maintain O balance. In this study, multi-step high-performance liquid chromatography (HPLC), combined with Mass Spectrometry (MS), was used to isolate and identify Cu/Zn-SOD from the serum of Pinctada fucata martensii (P. f. martensii) and was designated as PmECSOD. With a length of 1864 bp and an open reading frame (ORF) of 1422 bp, the cDNA encodes a 473 amino acid protein. The PmECSOD transcript was detected in multiple tissues by quantitative real-time PCR (qRT-PCR), with its highest expression level being in the gills. Additionally, the temporal expression of PmECSOD mRNA in the hemolymph was highest at 48 h after in vivo stimulation with Escherichia coli and Micrococcus luteus. The results from this study provide a valuable base for further exploration of molluscan innate immunity and immune response.
Topics: Animals; Pinctada; Superoxide Dismutase; Amino Acid Sequence; Immunity, Innate; Phylogeny; Gene Expression Profiling; Base Sequence; Sequence Alignment; Escherichia coli; DNA, Complementary; Micrococcus luteus; Gene Expression Regulation; RNA, Messenger
PubMed: 38701990
DOI: 10.1016/j.fsi.2024.109599 -
Journal of Materials Chemistry. B Sep 2021Superoxide dismutases (SODs) are a group of metalloenzymes that catalyze the dismutation of superoxide radicals (O˙) into hydrogen peroxide (HO) and oxygen (O). As the... (Review)
Review
Superoxide dismutases (SODs) are a group of metalloenzymes that catalyze the dismutation of superoxide radicals (O˙) into hydrogen peroxide (HO) and oxygen (O). As the first line of defense against reactive oxygen species (ROS)-mediated damage, SODs are expected to play an important role in the treatment of oxidative stress-related diseases. However, the clinical applications of SODs have been severely limited by their structural instability and high cost. Compared with natural enzymes, nanozymes, nanomaterials with enzyme-like activity, are more stable, and economical, can be easily modified and their activities can be adjusted. Due to their excellent characteristics, nanozymes have attracted widespread attention in recent years and are expected to become effective substitutes for natural enzymes in many application fields. Importantly, some nanozymes with SOD-like activity have been developed and proved to have a mitigating effect on diseases caused by oxidative stress. These studies on SOD-like nanozymes provide a feasible strategy for breaking through the dilemma of SOD clinical applications. However, at present, the specific catalytic mechanism of SOD-like nanozymes is still unclear, and many important issues need to be resolved. Although there are many comprehensive reviews to introduce the overall situation of the nanozyme field, the research on SOD-like nanozymes still lacks a systematic review. From the structure and mechanism of natural SOD enzymes to the structure and regulation of SOD-like nanozymes, and then to the measurement and application of nanozymes, this review systematically summarizes the recent progress in SOD-like nanozymes. The existing shortcomings and possible future research hotspots in the development of SOD-like nanozymes are summarized and prospected. We hope that this review would provide ideas and inspirations for further research on the catalytic mechanism and rational design of SOD-like nanozymes.
Topics: Animals; Antioxidants; Biocompatible Materials; Humans; Materials Testing; Superoxide Dismutase
PubMed: 34161407
DOI: 10.1039/d1tb00720c -
Journal of the College of Physicians... Jul 2021To compare superoxide dismutase 1 (SOD1) levels in diabetes mellitus patients with and without neuropathy.
OBJECTIVE
To compare superoxide dismutase 1 (SOD1) levels in diabetes mellitus patients with and without neuropathy.
STUDY DESIGN
Cross-sectional comparative study.
PLACE AND DURATION OF STUDY
Multidisciplinary Lab-1 of the Department of Biochemistry and Molecular Biology, Army Medical College, in cooperation with Pak-Emirates Military Hospital, Rawalpindi, Pakistan, from January 2020 to January 2021.
METHODOLOGY
Eighty-four subjects were enrolled through non-probability purposive sampling technique. They were further divided into three groups. Patients with diabetic neuropathy were labelled as the group Ι, and patients with diabetes mellitus without neuropathy were included in group ΙΙ. While group ΙΙΙ was comprised of healthy individuals and taken as control. Biochemical parameters included fasting blood glucose levels and HbA1c. Superoxide dismutase-1 levels were measured by enzyme-linked immunosorbent assay (ELISA). Data were evaluated by SPSS version 22.0 and presented in percentage and mean ± standard deviation (SD). Independent sample t-test and one-way ANOVA followed by the post-hoc Tukey test were used for group comparison.
RESULTS
Mean level of SOD1 was significantly higher in group Ι as compared to group ΙΙ and ΙΙΙ (p<0.001). Statistically significant difference was observed among groups Ι and ΙΙ (p=0.002), Ι and ΙΙΙ (p<0.001), and ΙΙ and ΙΙΙ (p=0.017). Mean levels of SOD1 were also significantly increased with poor glycemic control (p<0.001).
CONCLUSION
SOD1 level was considerably high in patients with diabetes mellitus with neuropathy in comparison to diabetics without neuropathy and healthy individuals. Key Words: Diabetes mellitus, Diabetic neuropathy, ELISA, Superoxide dismutase 1(SOD1).
Topics: Cross-Sectional Studies; Diabetes Mellitus, Type 2; Humans; Pakistan; Superoxide Dismutase; Superoxide Dismutase-1
PubMed: 34271773
DOI: 10.29271/jcpsp.2021.07.765 -
Free Radical Biology & Medicine Feb 2021Superoxide dismutase 3 (SOD3) is an extracellular protein with the capacity to convert superoxide into hydrogen peroxide, an important secondary messenger in redox...
Superoxide dismutase 3 (SOD3) is an extracellular protein with the capacity to convert superoxide into hydrogen peroxide, an important secondary messenger in redox regulation. To investigate the utility of zebrafish in functional studies of SOD3 and its relevance for redox regulation, we have characterized the zebrafish orthologues; Sod3a and Sod3b. Our analyses show that both recombinant Sod3a and Sod3b express SOD activity, however, only Sod3b is able to bind heparin. Furthermore, RT-PCR analyses reveal that sod3a and sod3b are expressed in zebrafish embryos and are present primarily in separate organs in adult zebrafish, suggesting distinct functions in vivo. Surprisingly, both RT-PCR and whole mount in situ hybridization showed specific expression of sod3b in skeletal tissue. To further investigate this observation, we compared femoral bone obtained from wild-type and SOD3 mice to determine whether a functional difference was apparent in healthy adult mice. Here we report, that bone from SOD3 mice is less mineralized and characterized by significant reduction of cortical and trabecular thickness in addition to reduced mechanical strength. These analyses show that SOD3 plays a hitherto unappreciated role in bone development and homeostasis.
Topics: Animals; Bone and Bones; Homeostasis; Mice; Mice, Knockout; Oxidation-Reduction; Superoxide Dismutase; Zebrafish
PubMed: 33476796
DOI: 10.1016/j.freeradbiomed.2021.01.027 -
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 -
Molecular Biology Reports Nov 2022Incomplete combustion of wood releases toxic chemicals. Exposure to these chemicals during charcoal production can modulate redox status of cellular system which may...
BACKGROUND
Incomplete combustion of wood releases toxic chemicals. Exposure to these chemicals during charcoal production can modulate redox status of cellular system which may further lead to genomic instability and of antioxidant enzymes. Genetic polymorphism may alter the functioning properties of these enzymes and modulate the response to oxidative stress.
METHODS
In this study, we analyzed the link between genetic polymorphism and enzyme activity for antioxidant enzymes: MnSOD and GPx-1 in charcoal workers and control population. This study included 77 charcoal workers and 79 demographically matched healthy control subjects. This association was studied using multiple linear regression, adjusted for confounding factors viz. age, consumption habits and exposure duration.
RESULTS
SOD activity was lower for TT genotype (3.47 ± 0.66; 5.92 ± 1.08) versus CC genotype (3.47 ± 0.66; 6.67 ± 1.60) in control and charcoal workers respectively. Significant lower GPx-1 activity was found in leu/leu genotype (7.25 ± 0.38; 3.59 ± 0.57) when compared to pro/pro genotype (7.78 ± 0.59; 4.28 ± 0.71) and pro/leu genotype (8.48 ± 0.34; 4.30 ± 0.76) in control population and charcoal workers respectively. A significant difference in the levels of 1-Hydroxypyrene (biomarker of exposure) and SOD and GPx-1 activity (biomarkers of oxidative stress) was evident in exposed group in comparison to the control one.
CONCLUSION
Collectively, our findings suggested that PAH influenced the mode of action of SOD and GPx-1 which were impacted by polymorphism in SOD and GPx-1 gene. Hence, polymorphism of MnSOD and GPx-1 genes were found to play a modulatory role in human susceptibility to oxidative damage induced by wood smoke in charcoal workers.
Topics: Humans; Glutathione Peroxidase; Antioxidants; Charcoal; Superoxide Dismutase; Polymorphism, Genetic; Oxidative Stress; Superoxide Dismutase-1; Biomarkers; Catalase
PubMed: 36074229
DOI: 10.1007/s11033-022-07779-2 -
Journal of Biomolecular Structure &... Sep 2022Superoxide dismutases (SODs) are regarded as important antioxidants for protecting cells against damage arising from oxidative stress. Much research is focused on...
Superoxide dismutases (SODs) are regarded as important antioxidants for protecting cells against damage arising from oxidative stress. Much research is focused on finding new chemicals with an ability to boost human SOD activity. In the research described herein a structure-based approach was used to identify new human Cu-Zn superoxide dismutase (SOD1) modulators based on previously reported plasmodium falciparum iron SOD inhibitors using induced fit docking and molecular dynamic (MD) protocols. The compound with the highest docking binding energy was selected for further structure simplification followed by structural similarity and MD in order to find a new activator/inhibitor scaffold of the SOD1 enzyme. According to the docking survey of the mentioned series, 1,4-bis(3-(1,4,8-trichloro-10Hphenothiazin-10-yl) propyl) piperazine (DS88) was the top scoring compound interacting with the SOD1 active site channel. Following structure simplification and similarity search, the most promising scaffold which is closely related to the phenothiazine antipsychotic class, was identified. Compared with the normal blood SOD1 activity, the percent of O production increased with trifluoperazine, while it decreased with the chlorpromazine. The molecular dynamic investigation shows that trifluoperazine exerts its SOD1 activating effect by stabilizing electrostatic loop while chlorpromazine employs SOD1 inhibition activity through repositioning of the electrostatic loop and increasing its distance from the catalytic metal site which diminished substrate specificity and catalytic activity of the SOD1 enzyme. The results identified the preferred region, orientation, and types of interaction for each activator or inhibitor compound.
Topics: Catalytic Domain; Chlorpromazine; Humans; Superoxide Dismutase; Superoxide Dismutase-1; Trifluoperazine
PubMed: 33663349
DOI: 10.1080/07391102.2021.1893819