THE ISOLATION OF PURE HYPOTAURINE FROM THE URINE OF RATS FED CYSTINE

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Experiments with D-cysteine in the rat.

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Chromatographic Evidence on the Occurrence of Thiotaurine in the Urine of Rats Fed with Cystine

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Electron paramagnetic resonance studies of cobalt-copper bovine superoxide dismutase.

Abstract 1. EPR spectra of cobalt-copper bovine superoxide dismutase at liquid nitrogen and liquid helium temperature show that the two metal centers are magnetically coupled. The temperature dependence of the spectra indicates that this coupling arises from an exchange interaction. 2. The EPR spectrum of the Co(II) of the enzyme can only be seen after reduction of the Cu(II), at very low temperature. It is typical of tetrahedral coordination which is distorted in a particular way. The EPR parameters are g⊥ ≃ 4, g|| ≃ 2, D = 11.5 cm-1. No feature indicating interaction between the two Co(II) centers is observed. 3. Anions such as CN- and N3- do not affect the EPR spectrum of Co(II) significantly, but only modify the spectrum of Cu(II). It is concluded that the Co(II) site (and presumably the native Zn(II) site) can be described as distorted tetrahedral, strongly spin-coupled to the Cu(II) and therefore very near to it, and noninteracting with the other Co(II) site and with solvent molecules

Kinetics of Electron Transfer between Azurin and Cytochrome 551 from Pseudomonas

Abstract The kinetics of electron transfer between the copper-containing protein azurin (Cu++/Cu+) and cytochrome 551 (Fe+++/Fe++) from Pseudomonas has been studied by rapid mixing methods. The reaction in both directions is fast; at low reagent concentrations (∼10-6 m) the apparent second order rate constant, at 20°, is about 3 x 106 m-1 sec-1 for the reaction Fe++ + Cu++ and 1.4 x 106 m-1 sec-1 for the reaction Fe+++ + Cu+. At high reagent concentrations the rates tend to reach a limiting value indicating that the reaction is not a simple second order process. The kinetics of the reactions of the reduced and oxidized forms of azurin and Pseudomonas cytochrome 551 with ferricyanide and dithionite has also been investigated. The rates of these reactions, at comparable reagent concentrations, are orders of magnitude lower than that between azurin and cytochrome 551

Catalase Takes Part in Rat Liver Mitochondria Oxidative Stress Defense

Highly purified rat liver mitochondria (RLM) when exposed to tert-butylhydroperoxide undergo matrix swelling, membrane potential collapse, and oxidation of glutathione and pyridine nucleotides, all events attributable to the induction of mitochondrial permeability transition. Instead, RLM, if treated with the same or higher amounts of H2O2 or tyramine, are insensitive or only partially sensitive, respectively, to mitochondrial permeability transition. In addition, the block of respiration by antimycin A added to RLM respiring in state 4 conditions, or the addition of H2O2, results in O2 generation, which is blocked by the catalase inhibitors aminotriazole or KCN. In this regard, H2O2 decomposition yields molecular oxygen in a 2:1 stoichiometry, consistent with a catalytic mechanism with a rate constant of 0.0346 s(-1). The rate of H2O2 consumption is not influenced by respiratory substrates, succinate or glutamate-malate, nor by N-ethylmaleimide, suggesting that cytochrome c oxidase and the glutathione-glutathione peroxidase system are not significantly involved in this process. Instead, H2O2 consumption is considerably inhibited by KCN or aminotriazole, indicating activity by a hemoprotein. All these observations are compatible with the presence of endogenous heme-containing catalase with an activity of 825 +/- 15 units, which contributes to mitochondrial protection against endogenous or exogenous H2O2. Mitochondrial catalase in liver most probably represents regulatory control of bioenergetic metabolism, but it may also be proposed for new therapeutic strategies against liver diseases. The constitutive presence of catalase inside mitochondria is demonstrated by several methodological approaches as follows: biochemical fractionating, proteinase K sensitivity, and immunogold electron microscopy on isolated RLM and whole rat liver tissue

Zymographic assay of plant diamine oxidase on entrapped peroxidase polyacrylamide gel electrophoresis. A study of stability to proteolysis

A zymographic assay of diamine oxidase (DAO, histaminase, EC 1.4.3.6), based on a coupled peroxidase reaction, and its behavior at proteolysis in simulated gastric and intestinal conditions, are described. The DAO activity from a vegetal extract of Lathyrus sativus seedlings was directly determined on sodium dodecyl sulfate polyacrylamide electrophoretic gels containing entrapped horseradish peroxidase, with putrescine as substrate of histaminase and ortho-phenylenediamine as co-substrate of peroxidase. The accumulation of azo-aniline, as peroxidase-catalyzed oxidation product, led to well-defined yellow-brown bands on gels, with intensities corresponding to the enzymatic activity of DAO. After image analysis of gels, a linear dependency of DAO content (Coomassie-stained protein bands) and of its enzymatic activity (zymographic bands) with the concentration of the vegetal extract was obtained. In simulated gastric conditions (pH 1.2, 37 °C), the DAO from the vegetal extract lost its enzymatic activity before 15 min of incubation, either in the presence or absence of pepsin. The protein pattern (Coomassie-stained) revealed that the DAO content from the vegetal extract was kept almost constant in the simulated intestinal fluid (containing pancreatin or not), with a slight diminution in the presence of pancreatic proteases. After 10 h of incubation at 37 °C, the DAO enzymatic activity from the vegetal extract was 44.7% in media without pancreatin and 13.6% in the presence of pancreatin, whereas the purified DAO retained only 4.65% of its initial enzymatic activity in the presence of pancreatin

Connection between amine oxidases and nitric oxide.

Conference on: Biogenic Amines 2005. Biological and Clinical Perspective, Giardini Naxos (ME)

Amine oxidases and tumors

The physiological function in living organisms of amine oxidases, is not completely established but is certainly related to the biogenic amines metabolism and therefore involved in essential processes such as the cell growth and differentiation. A correlation between degree of tumor malignancy and level of AO activity has been reported. The catalytic products of oxidative deamination of amines (hydrogen peroxide and aminoaldehyde) exert a cytotoxic effect and are considered cell growth inhibitors. The biogenic amines in same way could be considered in the cells as both poisons and protectors. A balance of oxidant and antioxidant enzymes appears to be very important in carcinogenesis and cell growth regulation. (C) 2003 Elsevier Inc. All rights reserved