Estimation of Disulfide Bonds Using Ellman’s Reagent

Aitken, Alastair; Learmonth, Michèle

doi:10.1007/978-1-59745-198-7_114

Cited by 29 publications

(31 citation statements)

References 4 publications

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“…Estimation of disulfide bonds amount in peptides bound to DNA was analyzed directly by using Ellman's reagent . The peptide/DNA complexes were prepared as described above at a concentration of 16 μ M for each peptide per sample.…”

Section: Methodsmentioning

confidence: 99%

Development of a receptor-targeted gene delivery system using CXCR4 ligand-conjugated cross-linking peptides

et al. 2014

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Section: Methodsmentioning

confidence: 99%

Development of a receptor-targeted gene delivery system using CXCR4 ligand-conjugated cross-linking peptides

et al. 2014

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“…. Total free sulfhydryl groups (SH) concentration was measured in the reaction with DTNB [35]. Briefly, myofibrillar proteins were dissolved in 20.0 mL urea-SDS buffer (8.0 M urea, 3% SDS and 0.1 M phosphate; pH 7.4) and left on the shaking bath at room temperature for 8 h. After that, 1 mL of the protein solution was incubated with 0.3 mL DTNB reagent (10 mM DTNB in 0.1 M phosphate buffer; pH 7.4) at room temperature for 15 min.…”

Section: Determination Of Sulfhydryl Groups Using 55′-dithiobis(2-nimentioning

confidence: 99%

Effect of Dietary Selenium on Protein and Lipid Oxidation and the Antioxidative Potential of Selected Chicken Culinary Parts during Frozen Storage

Korzeniowska

Króliczewska

Kopeć

2018

Journal of Chemistry

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The objective of the study was to evaluate the effects of inorganic and organic selenium in the diet of broiler chickens on the oxidative changes in the functional groups of proteins and total lipids, as well as the antioxidative potential of typical culinary parts fresh and frozen. Materials used in the nutrition study comprises one-day-old Flex broiler chickens randomly allocated to three dietary treatments: Control and SeN-fed diet enriched with 0.50 mg/kg of inorganic selenium (sodium selenite), and SeO-fed with diet containing 0.50 mg/kg of selenized yeast Yarrowia lipolytica. After slaughter, chicken carcasses were divided into the most typical culinary parts i.e., wings, outer and deep breasts, drumstick, thigh and back and analyzed in the fresh state and during frozen storage at −18°C until 90 days. The analyses undertaken during the study measured selenium concentration, CO, SH, and NH2 groups in the proteins, TBARS-expressing changes in the lipids, and antioxidative potential by ABTS, DPPH, and FRAP methods. The results of the study showed that the dietary selenium supplementation effectively increased the selenium concentration in all analyzed culinary parts of the chicken carcass, especially high in leg muscles. Selenium supplementation of the chicken diet significantly reduced the oxidative changes in the most important chemical reactive groups of the muscle myofibrillar proteins in all analyzed culinary parts. Both forms of selenium, organic and inorganic, were able to slow down the oxidation processes during first 30 days of the frozen storage of the meat. Longer storage could be only recommended for breast fillets, definitely not for the back part. The effect was much stronger in case of an organic selenium supplementation comparing to inorganic form of diet enrichment.

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“…Isolation of intracellular water soluble proteins was carried out on suspension cultures after 15 days of cultivation. Cells were grinded in liquid nitrogen and resuspended in 0.1 mol/L phosphate buffer, pH 8 [ 8 ] and centrifuged for 15 min at 15,000 × g at 4 °C to eliminate cellular residues. Supernatants containing diluted proteins were dialysed for 24 hours against a 1000-fold volume of phosphate buffer to eliminate low molecular weight thiol compounds.…”

Section: Methodsmentioning

confidence: 99%

“…Free thiols were determined by mixing equal volumes of protein sample and 10 × 10 −3 M 5,5′-dithiobis-(2-nitrobenzoic acid) (DTNB) in phosphate buffer (Ellman's reagent) and incubation for 15 min at room temperature, followed by absorbance measurement at 412 nm. [ 8 ] The concentration was calculated according to L-Cys standard curve. The total quantity of protein thiols was measured by the same protocol after preliminary reduction of protein and mixed disulphides with 20 × 10 −3 M dithiothreitol (DTT) and dialysis for three hours.…”

Section: Methodsmentioning

confidence: 99%

Changes in protein thiols in response to salt stress in embryogenic suspension cultures ofDactylis glomerataL.

Zagorchev

Terzieva

Stoichkova

et al. 2014

Biotechnology & Biotechnological Equipment

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The aim of the present study is to assess the rate of protein disulphide formation and the activity of NADPH-dependent thioredoxin and glutaredoxin systems, responsible for the reverse reduction of protein and mixed protein-glutathione disulphides, in embryogenic suspension cultures of Dactylis glomerata, subjected to salt stress. Two concentrations of NaCl previously established as enhancing (0.085 mol/L) and inhibiting (0.17 mol/L) somatic embryogenesis were used. The quantitative (by colour reaction with Ellman's reagent) and qualitative (by diagonal gel electrophoresis) analyses showed a significant increase in protein disulphide formation in salt-treated cultures compared to controls. The ratio of disulphides to free thiols is higher in 0.17 mol/L NaCl-treated cultures. The activity of the thioredoxin–thioredoxin reductase system has been increased accordingly in 0.085 mol/L NaCl-treated cultures but decreased at the higher salt concentration. The activity of glutaredoxins was also estimated, by using glutathionylated bovine serum albumin as substrate and following the decrease of NADPH absorbance at 340 nm in the presence of glutathione and glutathione reductase. Mild salt (0.085 mol/L NaCl) treated cultures again showed the highest activity compared to controls and 0.17 mol/L NaCl-treated cultures. Based on these observations it was suggested that salt treatment resulted in increased protein disulphide formation and thioredoxin and glutaredoxin systems are important regulators of this process, strongly involved in salt stress response. The highest activity at 0.085 mol/L NaCl may be also related to the regulatory mechanisms, involved in the potentiating of somatic embryogenesis at this salt concentration.

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Estimation of Disulfide Bonds Using Ellman’s Reagent

Cited by 29 publications

References 4 publications

Development of a receptor-targeted gene delivery system using CXCR4 ligand-conjugated cross-linking peptides

Development of a receptor-targeted gene delivery system using CXCR4 ligand-conjugated cross-linking peptides

Effect of Dietary Selenium on Protein and Lipid Oxidation and the Antioxidative Potential of Selected Chicken Culinary Parts during Frozen Storage

Changes in protein thiols in response to salt stress in embryogenic suspension cultures ofDactylis glomerataL.

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