scholarly journals Developmental regulation of rat lung Cu,Zn-superoxide dismutase

1987 ◽  
Vol 246 (3) ◽  
pp. 697-703 ◽  
Author(s):  
M A Hass ◽  
D Massaro
Keyword(s):  
Superoxide Dismutase ◽  
Half Life ◽  
Specific Activity ◽  
Developmental Regulation ◽  
Progressive Increase ◽  
Enzyme Synthesis ◽  
Enzyme Activation ◽  
Sod Activity ◽  
Adult Rats ◽  
Copper Chelation

In the present investigation we found that lung Cu, Zn-superoxide dismutase (SOD) activity (units/mg of DNA) increases steadily in the rat from birth to adulthood. The specific activity (units/micrograms of enzyme) of Cu, Zn-SOD was unchanged from birth to adulthood, excluding enzyme activation as a mechanism responsible for the increase in enzyme activity. Lung synthesis of Cu, Zn-SOD peaked at 1 day before birth and decreased thereafter to adult values. Calculations, based on rates of Cu, Zn-SOD synthesis and the tissue content of the enzyme, indicated that lung Cu, Zn-SOD activity increased during development owing to the rate of enzyme synthesis exceeding its rate of degradation by 5-10%. These calculations were supported by measurements of enzyme degradation in the neonatal (half-life, t1/2, = 12 h) and adult lung (t1/2 = greater than 100 h); the difference in half-life did not reflect the rates of overall protein degradation in the lung, since these rates were not different in lungs from neonatal and adult rats. We did not detect differences in the Mr or pI of Cu, Zn-SOD during development, but the susceptibility of the enzyme to inactivation by heat or copper chelation decreased with increasing age of the rats. We conclude that the progressive increase in activity of Cu, Zn-SOD is due to a rate of synthesis that exceeds degradation of the enzyme. The data also suggest that increased stabilization of enzyme conformation accounts for the greater half-life of the enzyme in lungs of adult compared with neonatal rats.

Alpha 1-adrenergic stimulation induces cardiac tolerance to hypoxia via induction and activation of Mn-SOD

1996 ◽  
Vol 271 (4) ◽  
pp. H1356-H1362 ◽  
Author(s):  
N. Yamashita ◽  
M. Nishida ◽  
S. Hoshida ◽  
J. Igarashi ◽  
M. Hori ◽  
...  
Keyword(s):  
Superoxide Dismutase ◽  
Creatine Kinase ◽  
Cardiac Myocytes ◽  
Total Protein ◽  
Manganese Superoxide Dismutase ◽  
Specific Activity ◽  
Adrenergic Stimulation ◽  
Sod Activity ◽  
Manganese Superoxide ◽  
Tolerance To Hypoxia

We examined whether or not alpha 1-adrenergic stimulation increases the tolerance of the heart to ischemia using a hypoxia-reoxygenation model of cardiac myocytes. After exposure to norepinephrine (NE; 0.2 microM) for 24 h, the manganese superoxide dismutase (Mn-SOD) content and activity in the cells were increased from 0.61 +/- 0.03 to 0.87 +/- 0.04 microgram/dish and 22 +/- 1 to 55 +/- 4 U/dish, respectively. The specific activity of Mn-SOD was also increased from 36 to 63 U/microgram Mn-SOD protein after the stimulation with NE. Prazosin (2 microM) abolished the increase in Mn-SOD activity (U/mg total protein). Creatine kinase (CK) release after hypoxia (PO2 7 mmHg; 3 h)-reoxygenation (1 h) from cells pretreated with NE in the presence of propranolol and yohimbine for 24 h was attenuated by 48% compared with that from cells without NE stimulation. When antisense oligodeoxyribonucleotides to Mn-SOD were added to myocyte cultures, the increase in Mn-SOD activity (U/mg total protein) and the attenuation of CK release after the addition of NE in the presence of propranolol and yohimbine were not observed. These results suggest that alpha 1-adrenergic stimulation increases the tolerance of myocytes to hypoxia through induction and activation of Mn-SOD.

scholarly journals Purification and partial characterization of superoxide dismutase from the thermophilic bacteria Thermothrix sp

2004 ◽  
Vol 69 (1) ◽  
pp. 9-16 ◽  
Author(s):  
Svetlana Seatovic ◽  
Ljubinka Gligic ◽  
Zeljka Radulovic ◽  
Ratko Jankov
Keyword(s):  
Molecular Weight ◽  
Superoxide Dismutase ◽  
Specific Activity ◽  
Thermophilic Bacteria ◽  
Noncovalent Interactions ◽  
Gel Filtration ◽  
Antioxidant Defense System ◽  
Exchange Chromatography ◽  
Gel Chromatography ◽  
Sod Activity

Superoxide dismutase (SOD; EC 1.15.1.1), a high molecular weight component of the antioxidant defense system, provided promising results in the treatment of oxidative damage. Thermothrix sp., isolated from thermal spa water in Serbia, showed high superoxide dismutase activity. The SOD, from cell free extract, was purified to homogenity by ammonium sulfate precipitation, Sephadex G 75 gel filtration chromatography and QAE Sephadex ion exchange chromatography. The specific activity of the purified enzyme was 9191 U/mg. The purified enzyme was analyzed and partially characterized. SOD was localized in polyacrylamide gel by activity staining, based on the reduction of nitroblue tetrazolium (NBT) by superoxide. The enzyme molecular weight determined by gel chromatography is 37 kD. According to SDS PAGE it is composed of two subunits of equal size, joined by noncovalent interactions. The isoelectric point, assessed by isoelectric focusing is 5.3. The optimum pH for enzyme activity was in the range of 8 to 10. The optimum temperature for SOD activity was 60 ?C. After one hour of incubation at 40, 50 and 60 ?C the SOD activity increases, but at 80 ?C, the SOD is denaturated. After 24 hours of incubation at 25 ?C SO Dactivity only slightly decreases.

scholarly journals Thermal stability of purified superoxide dismutase from liver of commercially valuable fish, Labeo rohita Exposed to Pb+Cr mixture

2021 ◽  
Vol 58 (04) ◽  
pp. 1191-1196
Author(s):  
Huma Naz
Keyword(s):  
Superoxide Dismutase ◽  
Specific Activity ◽  
Labeo Rohita ◽  
Maximum Activity ◽  
Exchange Chromatography ◽  
Sod Activity ◽  
Exposed Fish ◽  
Ammonium Sulphate Precipitation ◽  
Km Value ◽  
Increasing Temperature

In this experiment, effect of lead (Pb) + chromium (Cr) mixture on superoxide dismutase (SOD) in the liver of Labeo rohita at a concentration of 11.1 mgL-1 was observed. The ammonium sulphate precipitation and ion exchange chromatography techniques were successfully used to purify SOD. After purification, SOD activity of control and Pb+Cr treated fish was noted as 581.00 and 645.45 UmL-1, respectively while the specific activity was 1383.33 and 1613.62 Umg-1, respectively. The fold purification value of SOD was 2.75 and 2.45 for control and stressed fish, respectively. The recovery was calculated as 77.06 and 57.43% for control and stressed fish, respectively. The results of kinetic characterization showed that SOD form control and exposed fish had maximum activity at pH 6.5 and 7.0. Temperature also had a significant effect on activity of SOD. The SOD activity was measured maximum at 30°C for both control and Pb+Cr exposed fish. The Km value of liver SOD for control and Pb+Cr treated L. rohita was calculated as 1.48 and 0.62 mM, respectively. The value of Vmax for SOD from liver of control and Pb+Cr exposed fish was 1000 and 570 U mL-1, respectively. The enthalpy of denaturation (∆H*) for liver SOD from control and Pb+Cr exposed L. rohita was computed as 3.492 and 2.802 KJ mol-1 at 40°C, respectively and these values were dropped off with increasing the temperature until it remains 3.251 and 2.561 KJ mol-1 at 70°C, respectively. The free energy of thermal denaturation (ΔGº) of liver SOD was slightly increased with increasing temperature until 75°C which shows its resistance against heat. The values of ΔGº was observed as 58.03 and 57.95 KJ mol-1 for control and exposed fish at 40°C, respectively while the same was increased upto 62.37 and 62.00 KJ mol-1 at 70°C, respectively. It was concluded from negative value of ΔS* (entropy of inactivation) that the SOD is stable thermodynamically.

scholarly journals Molecular Cloning and Nucleotide Sequence of the Superoxide Dismutase Gene and Characterization of Its Product fromBacillus subtilis

1998 ◽  
Vol 180 (14) ◽  
pp. 3697-3703 ◽  
Author(s):  
Takashi Inaoka ◽  
Yoshinobu Matsumura ◽  
Tetsuaki Tsuchido
Keyword(s):  
Superoxide Dismutase ◽  
Stationary Phase ◽  
Specific Activity ◽  
Upstream Region ◽  
Gene Encoding ◽  
Sod Activity ◽  
Vegetative Cells ◽  
Reading Frame ◽  
Superoxide Dismutase Gene

ABSTRACT Bacillus subtilis was found to possess one detectable superoxide dismutase (Sod) in both vegetative cells and spores. The Sod activity in vegetative cells was maximal at stationary phase. Manganese was necessary to sustain Sod activity at stationary phase, but paraquat, a superoxide generator, did not induce the expression of Sod. The specific activity of purified Sod was approximately 2,600 U/mg of protein, and the enzyme was a homodimer protein with a molecular mass of approximately 25,000 per monomer. The gene encoding Sod, designatedsodA, was cloned by the combination of several PCR methods and the Southern hybridization method. DNA sequence analysis revealed the presence of one open reading frame consisting of 606 bp. Several putative promoter sites were located in the upstream region ofsodA. The deduced amino acid sequence showed high homology with other bacterial manganese Sods. Conserved regions in bacterial manganese Sod could also be seen. The phenotype of double mutantEscherichia coli sodA sodB, which could not grow in minimal medium without supplemental amino acids, was complemented by the expression of B. subtilis sodA.

Endotoxin increases lung Cu,Zn superoxide dismutase mRNA: O2 raises enzyme synthesis

1989 ◽  
Vol 257 (2) ◽  
pp. L61-L64 ◽  
Author(s):  
J. Iqbal ◽  
L. B. Clerch ◽  
M. A. Hass ◽  
L. Frank ◽  
D. Massaro
Keyword(s):  
Superoxide Dismutase ◽  
Enzyme Synthesis ◽  
Adult Rats ◽  
Air Breathing ◽  
Rat Lungs ◽  
In Vitro Exposure ◽  
Increased Activity

Administration of endotoxin to adult rats increases lung Cu,Zn superoxide activity after 72 h of exposure to greater than 95% O2. The increased activity is brought about mainly by a faster rate of Cu,Zn superoxide dismutase synthesis; rats treated with endotoxin but not exposed to hyperoxia do not exhibit these findings (Hass, Frank, and Massaro, J. Biol. Chem. 257: 9379-9383, 1982). We now report that 48 h after treatment of adult rats with endotoxin there was a decreased rate of Cu,Zn superoxide dismutase synthesis by lung slices from air- and O2- exposed rats, although, in both groups, the lung concentration of Cu,Zn superoxide dismutase mRNA was increased approximately 45%. Exposure of endotoxin-treated rats to greater than 95% O2 or air for an additional 24 h (72 h all told) resulted in continued elevation of Cu,Zn superoxide dismutase mRNA only in lungs of O2- exposed rats. In vitro exposure of lung slices from air-breathing saline- or endotoxin-treated rats to 95% O2 for 6 h led to an increased rate of Cu,Zn superoxide dismutase synthesis only in slices from endotoxin-treated rats. We conclude that endotoxin treatment leads to an increased concentration of Cu,Zn superoxide dismutase mRNA in rat lungs, but a sustained elevation of the mRNA, and its translation into an increased rate of Cu,Zn superoxide dismutase synthesis requires exposure of the lung to hyperoxia.

Pertussis toxin alters the concentration and turnover of manganese superoxide dismutase in rat lung

1996 ◽  
Vol 271 (6) ◽  
pp. L875-L879
Author(s):  
A. Berkovich ◽  
D. Massaro ◽  
L. B. Clerch
Keyword(s):  
Superoxide Dismutase ◽  
Pertussis Toxin ◽  
Manganese Superoxide Dismutase ◽  
Specific Activity ◽  
Oxygen Toxicity ◽  
Translational Efficiency ◽  
Sod Activity ◽  
Manganese Superoxide ◽  
Guanine Nucleotide Binding ◽  
Nucleotide Binding Proteins

Treatment of rats with pertussis toxin (PTX) decreases the activity of manganese superoxide dismutase (Mn-SOD) in the lung and results in oxygen toxicity in air (L. B. Clerch, G. Neithardt, U. Spencer, J. A. Melendez, G. D. Massaro, and D. Massaro. J. Clin. Invest. 93: 2482-2489, 1994). To examine aspects of the mechanism of the PTX-induced fall in Mn-SOD activity, we injected rats with PTX (50 micrograms/kg), killed the rats 72 h later, and measured the activity, concentration, specific activity, and turnover of Mn-SOD in the lung. Treatment with PTX caused an approximately 50% fall in Mn-SOD activity and Mn-SOD concentration but no change in Mn-SOD specific activity. PTX also caused an increase in Mn-SOD mRNA concentration, a fall in Mn-SOD synthesis, and an increase in the half-life of Mn-SOD and general proteins. We conclude the PTX-induced low concentration of Mn-SOD is due to a decrease of translational efficiency. We suggest that, under normoxic conditions, signal transduction via heterotrimeric guanine nucleotide binding proteins regulates the expression of Mn-SOD at the level of translation and Mn-SOD degradation.

A heat-stable Cu/Zn superoxide dismutase from the viscera of sardinelle (Sardinella aurita): purification and biochemical characterization

Biologia ◽  
2014 ◽  
Vol 69 (12) ◽  
Author(s):  
Hayet Ben Khaled ◽  
Naourez Ktari ◽  
Rayda Siala ◽  
Noomen Hmidet ◽  
Ahmed Bayoudh ◽  
...  
Keyword(s):  
Superoxide Dismutase ◽  
Biochemical Characterization ◽  
Specific Activity ◽  
Sequence Similarity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Native Enzyme ◽  
Sod Activity ◽  
Sardinella Aurita

AbstractSuperoxide dismutase (SOD) is an enzyme that protects against oxidative stress from superoxide radicals in living cells. This enzyme was extracted from sardinelle (Sardinella aurita) viscera, purified and characterized. The Cu/Zn-SOD was purified to homogeneity by the three-step procedure consisting of the heating at 65°C for 15 min, precipitation with ammonium sulphate (30–60%, w/v) and Sephadex G-100 gel filtration with a 7.17-fold increase in specific activity. The molecular weight of the native enzyme was estimated to be 40 kDa by G-125 gel filtration on HPLC column and that of the subunit mass, deduced by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, was 20 kDa. Thus the native enzyme appeared to be a homodimer. The optimum pH and temperature for the purified SOD activity were determined to be pH 7.0 and 40°C, respectively. It retained more than 85% of its initial activity after 1 h of incubation at 50°C. The enzyme had a broad stability pH range of 6.0–9.0. The N-terminal sequence of the purified enzyme was VLKAVCVLKGTGEVT. This sequence exhibited a high degree of sequence similarity with other fish Cu/Zn SODs.

The influence of UV-B radiation on the superoxide dismutase of maize, potato, sorghum, and wheat leaves

1999 ◽  
Vol 77 (1) ◽  
pp. 70-76 ◽  
Author(s):  
Isabel Santos ◽  
José Almeida ◽  
Roberto Salema
Keyword(s):  
Superoxide Dismutase ◽  
Specific Activity ◽  
Solanum Tuberosum L ◽  
Total Activity ◽  
Triticum Aestivum L ◽  
Sod Activity ◽  
Sod Isoforms ◽  
Pioneer Plants ◽  
Wheat Leaves ◽  
Inhibition Studies

The influence of UV-B radiation on superoxide dismutase (SOD, EC 1.15.1.1) with reference to the activity and types of isoenzymes was studied in leaves of C3 plants (potato, Solanum tuberosum L. cv. Désirée, and wheat, Triticum aestivum L. var. Almansor) and C4 plants (maize, Zea mays L. var. LG12, and sorghum, Sorghum Pr 8515 Pioneer). Plants were grown under controlled conditions in growth chambers with and without UV-B radiation. After 10 days of irradiation it was found that UV-B affected both the total activity of SOD and the number and amount of isoforms in all plants. The total specific activity of SOD increased significantly in wheat, maize, and potato, whereas a decline was induced in sorghum. Native gels revealed that UV-B caused preferential changes of the SOD isoforms in all plants used. Inhibition studies with cyanide and hydrogen peroxide showed that, in maize, UV-B radiation caused a large accumulation of one Mn-SOD and in contrast the level of the Cu,Zn-SOD isoforms decreased. In potato leaves, UV-B changed the pattern of SOD isoenzymes causing the appearance of one isoform and the disappearance of another. In wheat and sorghum the pattern of the isoenzymes was not altered, only their relative amounts. The rise in SOD activity in maize, potato, and wheat is possibly correlated with the tolerance of UV-B ascribed to these crops by different researchers and the sensitivity of sorghum to UV-B is possibly associated with the decrease in SOD activity.Key words: superoxide dismutase, UV-B, maize, potato, sorghum, wheat.

Secretion of extracellular superoxide dismutase in neonatal lungs

2000 ◽  
Vol 279 (5) ◽  
pp. L977-L984 ◽  
Author(s):  
Eva Nozik-Grayck ◽  
Christine S. Dieterle ◽  
Claude A. Piantadosi ◽  
Jan J. Enghild ◽  
Tim D. Oury
Keyword(s):  
Superoxide Dismutase ◽  
Disulfide Bonds ◽  
Developmental Regulation ◽  
Extracellular Superoxide Dismutase ◽  
Sod Activity ◽  
Rabbit Lung ◽  
Pulmonary Response ◽  
Extracellular Compartment ◽  
Neonatal Lung ◽  
Heparin Affinity

Extracellular superoxide dismutase (EC-SOD), the only known enzymatic scavenger of extracellular superoxide, may modulate reactions of nitric oxide (NO) in the lungs by preventing reactions between superoxide and NO. The regulation of EC-SOD has not been examined in developing lungs. We hypothesize that EC-SOD plays a pivotal role in the response to increased oxygen tension and NO in the neonatal lung. This study characterizes rabbit EC-SOD and investigates the developmental regulation of EC-SOD activity, protein expression, and localization. Purified rabbit EC-SOD was found to have several unique biochemical attributes distinct from EC-SOD in other species. Rabbit lung EC-SOD contains predominantly uncleaved subunits that do not form disulfide-linked dimers. The lack of intersubunit disulfide bonds may contribute to the decreased heparin affinity and lower EC-SOD content in rabbit lung. EC-SOD activity in rabbit lungs is low before birth and increases soon after gestation. In addition, the enzyme is localized intracellularly in preterm and term rabbit lungs. Secretion of active EC-SOD into the extracellular compartment increases with age. The changes in EC-SOD localization and activity have implications for the neonatal pulmonary response to oxidative stress and the biological activity of NO at birth.

CuZn superoxide dismutase activities from Fasciola hepatica

Parasitology ◽  
1998 ◽  
Vol 117 (6) ◽  
pp. 555-562 ◽  
Author(s):  
L. PIACENZA ◽  
R. RADI ◽  
F. GOÑI ◽  
C. CARMONA
Keyword(s):  
Molecular Weight ◽  
Superoxide Dismutase ◽  
Fasciola Hepatica ◽  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
Apparent Molecular Weight ◽  
Exchange Chromatography ◽  
Soluble Extract ◽  
Sod Activity

The levels of superoxide dismutase (SOD) were determined in detergent-soluble, somatic and excretion–secretion (E–S) preparations from adult Fasciola hepatica using the xanthine oxidase system and visualized in substrate gels. Compared to detergent-soluble and somatic extracts, E–S products showed the highest SOD activity (88 ·5 U/mg), indicating active release to the medium in which parasites were maintained. SOD specific activity was also detected at high levels in E–S products from 3-week-old and 5-week-old immature migrating flukes (25 and 143 U/mg, respectively). In all preparations except for the somatic extract, the activity was characterized as cyanide-sensitive CuZn SOD. Differences in SOD isoenzyme profiles between the extracts were observed in native polyacrylamide gel electrophoresis: the somatic and detergent-soluble extracts exhibited 1 band of activity while the E–S products from immature and adults flukes contained 2 and 3 migrating bands, respectively. SOD was purified from the detergent-soluble extract and E–S products of adult worms by a combination of ultrafiltration, gel filtration on Sephacryl S-200 HR and ion-exchange chromatography on QAE Sephadex A-50. The SOD from detergent-soluble extract showed, by SDS–PAGE analysis, 1 band of 16 kDa apparent molecular weight. The SOD from E–S products showed 2 bands of 16 and 60 kDa apparent molecular weight. N-terminal sequence analysis of the 16 kDa band from the detergent-soluble preparation showed some similarity with Schistosoma mansoni cytoplasmic SOD. These enzymes may have a potential role in the evasion of the oxidative burst killing mechanism by immune cells.

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