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The Journal of Biological Chemistry Jul 1997
Review
Topics: Animals; Bacterial Proteins; Escherichia coli Proteins; Humans; Mutation; Regulon; Superoxide Dismutase; Superoxides; Trans-Activators; Transcription Factors
PubMed: 9228011
DOI: 10.1074/jbc.272.30.18515 -
Brain : a Journal of Neurology Aug 2013Mutations in the gene superoxide dismutase 1 (SOD1) are causative for familial forms of the neurodegenerative disease amyotrophic lateral sclerosis. When the first SOD1... (Review)
Review
Mutations in the gene superoxide dismutase 1 (SOD1) are causative for familial forms of the neurodegenerative disease amyotrophic lateral sclerosis. When the first SOD1 mutations were identified they were postulated to give rise to amyotrophic lateral sclerosis through a loss of function mechanism, but experimental data soon showed that the disease arises from a--still unknown--toxic gain of function, and the possibility that loss of function plays a role in amyotrophic lateral sclerosis pathogenesis was abandoned. Although loss of function is not causative for amyotrophic lateral sclerosis, here we re-examine two decades of evidence regarding whether loss of function may play a modifying role in SOD1-amyotrophic lateral sclerosis. From analysing published data from patients with SOD1-amyotrophic lateral sclerosis, we find a marked loss of SOD1 enzyme activity arising from almost all mutations. We continue to examine functional data from all Sod1 knockout mice and we find obvious detrimental effects within the nervous system with, interestingly, some specificity for the motor system. Here, we bring together historical and recent experimental findings to conclude that there is a possibility that SOD1 loss of function may play a modifying role in amyotrophic lateral sclerosis. This likelihood has implications for some current therapies aimed at knocking down the level of mutant protein in patients with SOD1-amyotrophic lateral sclerosis. Finally, the wide-ranging phenotypes that result from loss of function indicate that SOD1 gene sequences should be screened in diseases other than amyotrophic lateral sclerosis.
Topics: Amyotrophic Lateral Sclerosis; Animals; Humans; Mice; Mice, Transgenic; Mutation; Phenotype; Superoxide Dismutase; Superoxide Dismutase-1
PubMed: 23687121
DOI: 10.1093/brain/awt097 -
PloS One 2013Aging in the world population has increased every year. Superoxide dismutase 2 (Mn-SOD or SOD2) protects against oxidative stress, a main factor influencing cellular...
Aging in the world population has increased every year. Superoxide dismutase 2 (Mn-SOD or SOD2) protects against oxidative stress, a main factor influencing cellular longevity. Polymorphisms in SOD2 have been associated with the development of neurodegenerative diseases, such as Alzheimer's and Parkinson's disease, as well as psychiatric disorders, such as schizophrenia, depression and bipolar disorder. In this study, all of the described natural variants (S10I, A16V, E66V, G76R, I82T and R156W) of SOD2 were subjected to in silico analysis using eight different algorithms: SNPeffect, PolyPhen-2, PhD-SNP, PMUT, SIFT, SNAP, SNPs&GO and nsSNPAnalyzer. This analysis revealed disparate results for a few of the algorithms. The results showed that, from at least one algorithm, each amino acid substitution appears to harmfully affect the protein. Structural theoretical models were created for variants through comparative modelling performed using the MHOLline server (which includes MODELLER and PROCHECK) and ab initio modelling, using the I-Tasser server. The predicted models were evaluated using TM-align, and the results show that the models were constructed with high accuracy. The RMSD values of the modelled mutants indicated likely pathogenicity for all missense mutations. Structural phylogenetic analysis using ConSurf revealed that human SOD2 is highly conserved. As a result, a human-curated database was generated that enables biologists and clinicians to explore SOD2 nsSNPs, including predictions of their effects and visualisation of the alignment of both the wild-type and mutant structures. The database is freely available at http://bioinfogroup.com/database and will be regularly updated.
Topics: Amino Acid Sequence; Computational Biology; Computer Simulation; Conserved Sequence; Databases, Genetic; Humans; Internet; Models, Molecular; Molecular Sequence Data; Mutation; Polymorphism, Single Nucleotide; Protein Conformation; Sequence Alignment; Superoxide Dismutase; User-Computer Interface
PubMed: 23785434
DOI: 10.1371/journal.pone.0065558 -
Current Biology : CB Apr 2002Cells deficient in repairing DNA double-strand breaks have an increased level of spontaneous chromosomal aberrations. Modulating the level of molecular oxygen and its... (Review)
Review
Cells deficient in repairing DNA double-strand breaks have an increased level of spontaneous chromosomal aberrations. Modulating the level of molecular oxygen and its reactive metabolites demonstrates that oxygen metabolism is a major source of genomic instability.
Topics: Animals; Antigens, Nuclear; Apoptosis; Chromosome Aberrations; DNA Damage; DNA Helicases; DNA Ligase ATP; DNA Ligases; DNA Repair; DNA-Binding Proteins; Ku Autoantigen; Neurons; Nuclear Proteins; Oxygen; Reactive Oxygen Species; Superoxide Dismutase; Superoxide Dismutase-1
PubMed: 11937046
DOI: 10.1016/s0960-9822(02)00788-1 -
Molecular Vision 2008To evaluate the age-induced changes in corneal endothelial morphology in mice lacking the cytosolic copper-zinc superoxide dismutase (SOD-1), the interstitial...
PURPOSE
To evaluate the age-induced changes in corneal endothelial morphology in mice lacking the cytosolic copper-zinc superoxide dismutase (SOD-1), the interstitial extracellular superoxide dismutase (SOD-3), or both of these SOD isoenzymes.
METHODS
The central corneal endothelial morphologies of old C57BL-6J wild type (n=19), SOD-1 null (n=16), SOD-3 null (n=15), and SOD1/3 null (n=11) mice were evaluated using alizarin red staining and light microscope photographs. For comparison, young endothelia from the same genotypes were evaluated similarly. The levels of corneal reactive oxygen species and nitrogen species in all four genotypes were quantified using fluorimetry with 2',7'-dichlorodihydrofluorescein diacetate and OxyBURST.
RESULTS
In accordance with our previous findings, the mean corneal endothelial cell area was larger in the SOD-3 null genotype than in the wild type mice. The SOD-1/3 null genotype had similar cell sizes as the SOD-3 null mice but had a more irregular morphology at an older age. Apparently, these irregularities develop with time as they are not seen in young animals. The SOD-1 null mice did not differ from the wild type mice in corneal endothelial morphology. Elevated levels of reactive oxygen species were seen in SOD-1 null and SOD-3 null corneas, and elevated superoxide levels were seen in all three knockout genotypes.
CONCLUSIONS
The increased spontaneous age-related enlargement of corneal endothelial cells seen in the absence of SOD-3 is associated with a more irregular cell pattern when combined with a lack of SOD-1. This indicates more cellular movements and ongoing repair in the SOD-1/3 null genotype and possibly a more vulnerable corneal endothelium. SOD-3 and SOD-1 appear to have functions in preserving corneal endothelial integrity in aging.
Topics: Aging; Animals; Endothelium, Corneal; Mice; Mice, Inbred C57BL; Reactive Nitrogen Species; Reactive Oxygen Species; Superoxide Dismutase; Superoxide Dismutase-1
PubMed: 18989385
DOI: No ID Found -
BMB Reports Feb 2015Cells express several antioxidant enzymes to scavenge reactive oxygen species (ROS) responsible for oxidative damages and various human diseases. Therefore, antioxidant...
Cells express several antioxidant enzymes to scavenge reactive oxygen species (ROS) responsible for oxidative damages and various human diseases. Therefore, antioxidant enzymes are considered biomedicine candidates. Among them, extracellular superoxide dismutase (SOD3) had showed prominent efficacy against asthma and inflammation. Despite its advantages as a biomedicine, the difficulty in obtaining large quantity of active recombinant human SOD3 (rhSOD3) has limited its clinical applications. We found that a significant fraction of overexpressed rhSOD3 was composed of the inactive apo-enzyme and its potency against inflammation depended on the rate of metal incorporation. Also, purified rhSOD3 was unstable and lost its activity very quickly. Here, we suggest an ideal preparative method to express, purify, and store highly active rhSOD3. The enzymatic activity of rhSOD3 was maximized by incorporating metal ions into rhSOD3 after purification. Also, albumin or polyethylene glycol prevented rapid inactivation or degradation of rhSOD3 during preparative procedures and long-term storage.
Topics: Copper; HEK293 Cells; Humans; Polyethylene Glycols; Protein Stability; Reactive Oxygen Species; Recombinant Fusion Proteins; Superoxide Dismutase; Zinc
PubMed: 24856831
DOI: 10.5483/bmbrep.2015.48.2.093 -
The Journal of Biological Chemistry Dec 2022The heterodimerization of WT Cu, Zn superoxide dismutase-1 (SOD1), and mutant SOD1 might be a critical step in the pathogenesis of SOD1-linked amyotrophic lateral...
The heterodimerization of WT Cu, Zn superoxide dismutase-1 (SOD1), and mutant SOD1 might be a critical step in the pathogenesis of SOD1-linked amyotrophic lateral sclerosis (ALS). Rates and free energies of heterodimerization (ΔG) between WT and ALS-mutant SOD1 in mismatched metalation states-where one subunit is metalated and the other is not-have been difficult to obtain. Consequently, the hypothesis that under-metalated SOD1 might trigger misfolding of metalated SOD1 by "stealing" metal ions remains untested. This study used capillary zone electrophoresis and mass spectrometry to track heterodimerization and metal transfer between WT SOD1, ALS-variant SOD1 (E100K, E100G, D90A), and triply deamidated SOD1 (modeled with N26D/N131D/N139D substitutions). We determined that rates of subunit exchange between apo dimers and metalated dimers-expressed as time to reach 30% heterodimer-ranged from t = 67.75 ± 9.08 to 338.53 ± 26.95 min; free energies of heterodimerization ranged from ΔG = -1.21 ± 0.31 to -3.06 ± 0.12 kJ/mol. Rates and ΔG values of partially metalated heterodimers were more similar to those of fully metalated heterodimers than apo heterodimers, and largely independent of which subunit (mutant or WT) was metal-replete or metal-free. Mass spectrometry and capillary electrophoresis demonstrated that mutant or WT 4Zn-SOD1 could transfer up to two equivalents of Zn to mutant or WT apo-SOD1 (at rates faster than the rate of heterodimerization). This result suggests that zinc-replete SOD1 can function as a chaperone to deliver Zn to apo-SOD1, and that WT apo-SOD1 might increase the toxicity of mutant SOD1 by stealing its Zn.
Topics: Humans; Superoxide Dismutase-1; Amyotrophic Lateral Sclerosis; Superoxide Dismutase; Metals; Zinc; Mutation
PubMed: 36265587
DOI: 10.1016/j.jbc.2022.102610 -
The FEBS Journal Nov 2006Human manganese superoxide dismutase is a mitochondrial metalloenzyme that is involved in protecting aerobic organisms against superoxide toxicity, and has been...
Human manganese superoxide dismutase is a mitochondrial metalloenzyme that is involved in protecting aerobic organisms against superoxide toxicity, and has been implicated in slowing tumor growth. Unfortunately, this enzyme exhibits strong product inhibition, which limits its potential biomedical applications. Previous efforts to alleviate human manganese superoxide dismutase product inhibition utilized rational protein design and site-directed mutagenesis. These efforts led to variants of human manganese superoxide dismutase at residue 143 with dramatically reduced product inhibition, but also reduced catalytic activity and efficiency. Here, we report the use of a directed evolution approach to engineer two variants of the Q143A human manganese superoxide dismutase mutant enzyme with improved catalytic activity and efficiency. Two separate activity-restoring mutations were found--C140S and N73S--that increase the catalytic efficiency of the parent Q143A human manganese superoxide dismutase enzyme by up to five-fold while maintaining low product inhibition. Interestingly, C140S is a context-dependent mutation, and the C140S-Q143A human manganese superoxide dismutase did not follow Michaelis-Menten kinetics. The re-engineered human manganese superoxide dismutase mutants should be useful for biomedical applications, and our kinetic and structural studies also provide new insights into the structure-function relationships of human manganese superoxide dismutase.
Topics: Catalysis; Humans; Kinetics; Models, Molecular; Mutation; Recombinant Proteins; Superoxide Dismutase
PubMed: 16999822
DOI: 10.1111/j.1742-4658.2006.05484.x -
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 -
International Journal of Molecular... Jan 2011A superoxide dismutase gene from thermotolerant Bacillus sp. MHS47 (MnSOD47) was cloned, sequenced, and expressed. The gene has an open reading frame of 612 bp,...
A superoxide dismutase gene from thermotolerant Bacillus sp. MHS47 (MnSOD47) was cloned, sequenced, and expressed. The gene has an open reading frame of 612 bp, corresponding to 203 deduced amino acids, with high homology to the amino acid sequences of B. thuringiensis (accession no. EEN01322), B. anthracis (accession no. NP_846724), B. cereus (accession no. ZP_04187911), B. weihenstephanensis (accession no. YP_001646918), and B. pseudomycoides. The conserved manganese-binding sites (H28, H83, D165, and H169) show that MnSOD47 has the specific characteristics of the manganese superoxide dismutase (MnSOD) enzymes. MnSOD47 expressed an enzyme with a molecular weight of approximately 22.65 kDa and a specific activity of 3537.75 U/mg. The enzyme is active in the pH range 7-8.5, with an optimum pH of 7.5, and at temperatures in the range 30-45 °C, with an optimum temperature of 37 °C. Tests of inhibitors and metal ions indicated that the enzyme activity is inhibited by sodium azide, but not by hydrogen peroxide or potassium cyanide. These data should benefit future studies of MnSODs in other microorganisms and the biotechnological production of MnSOD47, and could also be used to develop a biosensor for the detection of antioxidants and free radical activity. In the future, this basic knowledge could be applicable to the detection of cancer risks in humans and therapeutic treatments.
Topics: Bacillus; Bacterial Proteins; Enzyme Stability; Hydrogen-Ion Concentration; Superoxide Dismutase; Temperature
PubMed: 21340017
DOI: 10.3390/ijms12010844