scholarly journals Reactivity of hydroxyl and hydroxyl-like radicals discriminated by release of thiobarbituric acid-reactive material from deoxy sugars, nucleosides and benzoate

1984 ◽  
Vol 224 (3) ◽  
pp. 761-767 ◽  
Author(s):  
J M C Gutteridge
Keyword(s):  
Binding Sites ◽  
Thiobarbituric Acid ◽  
Iron Ions ◽  
Reactive Material ◽  
Reaction Systems ◽  
Ferric Salt ◽  
Iron Salts ◽  
A Site ◽  
Generating System ◽  
Deoxy Sugars

Hydroxyl radicals (OH.) can be formed in aqueous solution by a superoxide (O2.−)–generating system in the presence of a ferric salt or in a reaction independent of O2.- by the direct addition of a ferrous salt. OH. damage was detected in the present work by the release of thiobarbituric acid-reactive material from deoxy sugars, nucleosides and benzoate. The carbohydrates deoxyribose, deoxygalactose and deoxyglucose were substantially degraded by the iron(II) salt and the iron(III) salt in the presence of an O2.− –generating system, whereas deoxyinosine, deoxyadenosine and benzoate were not. Addition of EDTA to the reaction systems producing radicals greatly enhanced damage to deoxyribose, deoxyinosine, deoxyadenosine and benzoate, but decreased damage to deoxygalactose and deoxyglucose. Further, OH. scavengers were effective inhibitors only when EDTA was present. Inhibition by catalase and desferrioxamine confirmed that H2O2 and iron salts were essential for these reactions. The results suggest that, in the absence of EDTA, iron ions bind to the carbohydrate detector molecules and bring about a site-specific reaction on the molecule. This reaction is poorly inhibited by most OH. scavengers, but is strongly inhibited by scavengers such as mannitol, glucose and thiourea, which can themselves bind iron ions, albeit weakly. In the presence of EDTA, however, iron is removed from these binding sites to produce OH. in ‘free’ solution. These can be readily intercepted by the addition of OH. scavengers.

scholarly journals Hydroxyl-radical-induced iron-catalysed degradation of 2-deoxyribose. Quantitative determination of malondialdehyde

1988 ◽  
Vol 252 (3) ◽  
pp. 649-653 ◽  
Author(s):  
K H Cheeseman ◽  
A Beavis ◽  
H Esterbauer
Keyword(s):  
Quantitative Determination ◽  
Hydroxyl Radical ◽  
Hydroxyl Radicals ◽  
Degradation Product ◽  
Thiobarbituric Acid ◽  
Iron Ions ◽  
Reactive Material ◽  
Gamma Radiolysis ◽  
Deoxyribose Degradation

The degradation of 2-deoxyribose to thiobarbituric acid-reactive material was investigated with two hydroxyl-radical-generating systems: (i) a defined gamma-radiolysis method and (ii) incubation with FeSO4 in phosphate buffer. In each case the thiobarbituric acid-reactive material can be accounted for by malondialdehyde, as measured by an h.p.l.c. method for free malondialdehyde. In the radiolysis system there is a large post-irradiation increase in free malondialdehyde if iron ions are added to the samples. It is proposed that this is due to iron ions catalysing the formation of hydroxyl radicals from radiolytically generated H2O2 as well as stimulating the breakdown of an intermediate deoxyribose degradation product. A mechanism for the formation of malondialdehyde during deoxyribose degradation is proposed.

Lipid peroxidation in rheumatoid arthritis: Thiobarbituric acid-reactive material and catalytic iron salts in synovial fluid from rheumatoid patients

1984 ◽  
Vol 66 (6) ◽  
pp. 691-695 ◽  
Author(s):  
David Rowley ◽  
John M. C. Gutteridge ◽  
David Blake ◽  
Margaret Farrs ◽  
Barry Halliwell
Keyword(s):  
Rheumatoid Arthritis ◽  
Lipid Peroxidation ◽  
Synovial Fluid ◽  
Knee Joint ◽  
Thiobarbituric Acid ◽  
Radical Reactions ◽  
Local Inflammation ◽  
Reactive Material ◽  
Iron Salts

1. Thiobarbituric acid (TBA)-reactive material is present in serum and knee joint synovial fluid from rheumatoid patients, consistent with lipid peroxidation occurring in vivo. 2. The amount of TBA-reactive material in synovial fluid correlates with the concentration of iron salts present as determined by the bleomycin method, presumably because iron is an important catalyst of radical reactions in vivo. 3. There appear to be significant correlations between the contents of TBA-reactive material and bleomycindetectable iron in synovial fluid and the activity of rheumatoid arthritis as assessed with a clinical index of local inflammation and with various laboratory parameters.

scholarly journals Research on the effects of ferric salt added into aeration tank for phosphorus removal on biochemical system

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Xie Jingliang ◽  
Sun Yuanjie ◽  
Peng Anran ◽  
Faris Kateb ◽  
Hooreya Mohamed Ahmed Aldeeb
Keyword(s):  
Phosphorus Removal ◽  
Quality Parameters ◽  
Stable Operation ◽  
Aeration Tank ◽  
Iron Ions ◽  
Ferrous Chloride ◽  
Iron Salt ◽  
Biochemical System ◽  
Ferric Salt ◽  
Iron Salts

Abstract In this paper, two iron salts, ferrous chloride (FeCl2) and ferric chloride (FeCl3), are directly added into an aeration tank for phosphorus removal, and their effects on the biochemical system are studied; the water quality parameters such as pH and alkalinity are also investigated. The extent of influence of the added iron salts on the pH and alkalinity of aerated solutions is demonstrated to be FeCl3 > FeCl2. When the dosage of iron ions is 20 mg/L, the decrease in pH and alkalinity caused by FeCl3 is 0.5 and 65 mg/L, which is higher than FeCl2 by 2% and 26%. The initial phosphorus removal effect of FeCl2 is worse than that of FeCl3, but after continued aeration and oxidation, the phosphorus removal effect of FeCl2 can be improved; however, the final phosphorus removal effect is basically the same as that of FeCl3 added directly. The results show that FeCl2 is preferred when iron salt is added directly into the aeration tank to remove phosphorus. The proposed scheme can reduce the effect of iron salts on the alkalinity of the biochemical system on the premise of ensuring the phosphorus removal effect of the system, and is conducive to ensuring the stable operation of the biochemical system.

scholarly journals Effect of ferritin-containing fractions with different iron loading on lipid peroxidation

1983 ◽  
Vol 209 (2) ◽  
pp. 557-560 ◽  
Author(s):  
J M C Gutteridge ◽  
B Halliwell ◽  
A Treffry ◽  
P M Harrison ◽  
D Blake
Keyword(s):  
Lipid Peroxidation ◽  
Thiobarbituric Acid ◽  
Bovine Brain ◽  
Iron Loading ◽  
Reactive Material ◽  
Phospholipid Liposomes ◽  
Iron Salts ◽  
Brain Phospholipid ◽  
Equivalent Concentration ◽  
Iron Concentrations

Ferritin-containing fractions with different degrees of iron loading were prepared. All ferritin fractions stimulated the peroxidation of bovine brain phospholipid liposomes, as measured by the formation of thiobarbituric acid-reactive material. This stimulation was increased in the presence of ascorbate. Iron salts of equivalent concentration to those of the ferritin fractions were more stimulatory to lipid peroxidation at the higher iron concentrations. None of the fractions inhibited ascorbate-dependent peroxidation in the presence of added iron salts.

scholarly journals Crystal structures of Ca2+–calmodulin bound to NaV C-terminal regions suggest role for EF-hand domain in binding and inactivation

2019 ◽  
Vol 116 (22) ◽  
pp. 10763-10772 ◽  
Author(s):  
Bernd R. Gardill ◽  
Ricardo E. Rivera-Acevedo ◽  
Ching-Chieh Tung ◽  
Filip Van Petegem
Keyword(s):  
Crystal Structure ◽  
Binding Sites ◽  
Binding Mode ◽  
Conformational Switch ◽  
Channel Inactivation ◽  
Dependent Inactivation ◽  
Ef Hand ◽  
Inherited Arrhythmia ◽  
A Site

Voltage-gated sodium (NaV) and calcium channels (CaV) form targets for calmodulin (CaM), which affects channel inactivation properties. A major interaction site for CaM resides in the C-terminal (CT) region, consisting of an IQ domain downstream of an EF-hand domain. We present a crystal structure of fully Ca2+-occupied CaM, bound to the CT of NaV1.5. The structure shows that the C-terminal lobe binds to a site ∼90° rotated relative to a previous site reported for an apoCaM complex with the NaV1.5 CT and for ternary complexes containing fibroblast growth factor homologous factors (FHF). We show that the binding of FHFs forces the EF-hand domain in a conformation that does not allow binding of the Ca2+-occupied C-lobe of CaM. These observations highlight the central role of the EF-hand domain in modulating the binding mode of CaM. The binding sites for Ca2+-free and Ca2+-occupied CaM contain targets for mutations linked to long-QT syndrome, a type of inherited arrhythmia. The related NaV1.4 channel has been shown to undergo Ca2+-dependent inactivation (CDI) akin to CaVs. We present a crystal structure of Ca2+/CaM bound to the NaV1.4 IQ domain, which shows a binding mode that would clash with the EF-hand domain. We postulate the relative reorientation of the EF-hand domain and the IQ domain as a possible conformational switch that underlies CDI.

scholarly journals Characterization of the two 5-aminolaevulinic acid binding sites, the A- and P-sites, of 5-aminolaevulinic acid dehydratase from Escherichia coli

1995 ◽  
Vol 305 (1) ◽  
pp. 151-158 ◽  
Author(s):  
P Spencer ◽  
P M Jordan
Keyword(s):  
Schiff Base ◽  
Binding Sites ◽  
Metal Ion ◽  
Substrate Binding ◽  
Aminolaevulinic Acid ◽  
Acid Dehydratase ◽  
Carboxylate Groups ◽  
Catalytic Metal ◽  
A Site ◽  
Aminolaevulinic Acid Dehydratase

Experiments are described in which the individual properties of the two 5-aminolaevulinic acid (ALA) binding sites, the A-site and the P-site, of 5-aminolaevulinic acid dehydratase (ALAD) have been investigated. The ALA binding affinity at the A-site is greatly enhanced (at least 10-fold) on the binding of the catalytic metal ion (bound at the alpha-site). The nature of the catalytic metal ion, Mg2+ or Zn2+, also gave major variations in the substrate Km, P-site affinity for ALA, the effect of potassium and phosphate ions and the pH-dependence of substrate binding. Modification of the P-site by reaction of the enzyme-substrate Schiff base with NaBH4 and analysis of the reduced adduct by electro-spray mass spectrometry indicated a maximum of 1 mol of substrate incorporated/mol of subunit, correlating with a linear loss of enzyme activity. The reduced Schiff-base adduct was used to investigate substrate binding at the A-site by using rate-of-dialysis analysis. The affinity for ALA at the A-site of Mg alpha Zn beta ALAD was found to determine the Km for the reaction and was pH-dependent, with its affinity increasing from 1 mM at pH 6 to 70 microM at pH 8.5. The affinity of ALA at the P-site of Zn alpha An beta ALAD is proposed to limit the Km at pH values above 7, since the measured Kd for ALA at the A-site in 45 microM Tris, pH 8, was well below the observed Km (600 microM) under the same conditions. The amino group of the ALA molecule bound at the P-site was identified as a critical binding component for the A-site, explaining why ALA binding to ALAD is ordered, with the P-site ALA binding first. Structural requirements for ALA binding at the A- and P-sites have been identified: the P-site requires the carbonyl and carboxylate groups, whereas the A-site requires the amino, carbonyl and carboxylate groups of the substrate.

scholarly journals Specific Binding of Rubidium in Chlorella

1962 ◽  
Vol 45 (5) ◽  
pp. 959-977 ◽  
Author(s):  
Dan Cohen
Keyword(s):  
Active Transport ◽  
Binding Sites ◽  
Specific Binding ◽  
High Concentration ◽  
External Concentration ◽  
Specific Binding Sites ◽  
Dense Suspension ◽  
Concentration Of Ions ◽  
The Individual ◽  
A Site

Specific binding sites for potassium, which may be components of the carriers for active transport for K in Chlorella, were characterized by their capacity to bind rubidium. A dense suspension was allowed to take up Rb86 from a low concentration of Rb86 and a high concentration of ions which saturate non-specific sites. The amount bound was derived from the increase in the external concentration of Rb86 following addition of excess potassium. The sites were heterogeneous. The average affinity of Rb and various other ions for the sites was determined by plotting the degree of displacement of Rb86 against log molar concentration of the individual ions. Interpolation gave the concentration for 50 per cent displacement of Rb, which is inversely related to affinity. The order of affinity was not changed when the cells were frozen, or boiled either in water or in 70 per cent ethanol. The affinity is maximal for ions with a crystalline radius of 1.3 to 1.5 A and a high polarizability, and is not related to the hydrated radius or valency. It is suggested that binding groups in a site are rigidly arranged, the irregular space between them being 2.6 to 3.0 A across, so that affinity is high for ions of this diameter and high polarizability.

Estimation of 3-Deoxy Sugars by Means of the Malonaldehyde–Thiobarbituric Acid Reaction

Nature ◽  
1960 ◽  
Vol 186 (4719) ◽  
pp. 155-156 ◽  
Author(s):  
MORRIS A. CYNKIN ◽  
GILBERT ASHWELL
Keyword(s):  
Thiobarbituric Acid ◽  
Acid Reaction ◽  
Deoxy Sugars

Defining a second epitope for heparin-induced thrombocytopenia/thrombosis antibodies using KKO, a murine HIT-like monoclonal antibody

Blood ◽  
2002 ◽  
Vol 99 (4) ◽  
pp. 1230-1236 ◽  
Author(s):  
Zhong Q. Li ◽  
Weiyi Liu ◽  
Kwang S. Park ◽  
Brue S. Sachais ◽  
Gowthani M. Arepally ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Binding Site ◽  
Binding Sites ◽  
Heparin Therapy ◽  
Common Complication ◽  
Platelet Factor ◽  
Heparin Induced Thrombocytopenia ◽  
The Third ◽  
N Terminus ◽  
A Site

Heparin-induced thrombocytopenia/thrombosis (HIT/T) is a common complication of heparin therapy that is caused by antibodies to platelet factor 4 (PF4) complexed with heparin. The immune response is polyclonal and polyspecific, ie, more than one neoepitope on PF4 is recognized by HIT/T antibodies. One such epitope has been previously identified; it involves the domain between the third and fourth cysteine residues in PF4 (site 1). However, the binding sites for other HIT/T antibodies remain to be defined. To explore this issue, the binding site of KKO, an HIT/T-like murine monoclonal antibody, was defined. KKO shares a binding site with many HIT/T antibodies on PF4/heparin, but does not bind to site 1 or recognize mouse PF4/heparin. Therefore, the binding of KKO to a series of mouse/human PF4 chimeras complexed with heparin was examined. KKO recognizes a site that requires both the N terminus of PF4 and Pro34, which immediately precedes the third cysteine. Both regions lie on the surface of the PF4 tetramer in sufficient proximity (within 0.74 nm) to form a contiguous antigenic determinant. The 10 of 14 HIT/T sera that require the N terminus of PF4 for antigen recognition also require Pro34 to bind. This epitope, termed site 2, lies adjacent to site 1 in the crystal structure of the PF4 tetramer. Yet sites 1 and 2 can be recognized by distinct populations of antibodies. These studies further help to define a portion of the PF4 tetramer to which self-reactive antibodies develop in patients exposed to heparin.

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