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Iron Binding (iron + binding)
Terms modified by Iron Binding Selected AbstractsDynamics, stability and iron-binding activity of frataxin clinical mutantsFEBS JOURNAL, Issue 14 2008Ana R. Correia Friedreich's ataxia results from a deficiency in the mitochondrial protein frataxin, which carries single point mutations in some patients. In the present study, we analysed the consequences of different disease-related mutations in vitro on the stability and dynamics of human frataxin. Two of the mutations, G130V and D122Y, were investigated for the first time. Analysis by CD spectroscopy demonstrated a substantial decrease in the thermodynamic stability of the variants during chemical and thermal unfolding (wild-type > W155R > I154F > D122Y > G130V), which was reversible in all cases. Protein dynamics was studied in detail and revealed that the mutants have distinct propensities towards aggregation. It was observed that the mutants have increased correlation times and different relative ratios between soluble and insoluble/aggregated protein. NMR showed that the clinical mutants retained a compact and relatively rigid globular core despite their decreased stabilities. Limited proteolysis assays coupled with LC-MS allowed the identification of particularly flexible regions in the mutants; interestingly, these regions included those involved in iron-binding. In agreement, the iron metallochaperone activity of the Friedreich's ataxia mutants was affected: some mutants precipitate upon iron binding (I154F and W155R) and others have a lower binding stoichiometry (G130V and D122Y). Our results suggest that, in heterozygous patients, the development of Friedreich's ataxia may result from a combination of reduced efficiency of protein folding and accelerated degradation in vivo, leading to lower than normal concentrations of frataxin. This hypothesis also suggests that, although quite different from other neurodegenerative diseases involving toxic aggregation, Friedreich's ataxia could also be linked to a process of protein misfolding due to specific destabilization of frataxin. [source] Computational study of the solvation of protoporphyrin IX and its Fe2+ complexINTERNATIONAL JOURNAL OF QUANTUM CHEMISTRY, Issue 13 2008Teobaldo Cuya Guizado Abstract Molecular dynamics (MD) simulations of a well known hydrophobic structure, the heme (ferroprotoporphyrin IX) and its precursor in the heme synthesis, protoporphyrin IX (PPIX) are presented. The objective of the present study is to determine the stability of both structures in an aqueous medium, as well as the structure-solvent relation, hydration shells, and discuss their implications for biological processes. The density functional theory (DFT) is used for the electronic and structural characterization of both PPIX and its Fe2+ complex. A classical approach based on the Gromacs package is used for the MD. The radial distribution function g(r) is used to examine the allocation of water molecules around different regions of the porphyrins. The calculations demonstrate the heterogeneous character of the porphyrins with respect to the affinity with water molecules, the general hydrophobic character of the porphyrin ring bonded or not to the ion Fe, the hydrophilic character of the carboxylic oxygen that is unchanged upon iron binding, and the low hydrophilicity of Fe2+ in the heme. © 2008 Wiley Periodicals, Inc. Int J Quantum Chem, 2008 [source] Evaluation of Garam Masala Spices and Phosphates as Antioxidants in Cooked Ground BeefJOURNAL OF FOOD SCIENCE, Issue 5 2006Mihir N. Vasavada ABSTRACT:, This study determined antioxidant effects and sensory attributes of individual ingredients (black pepper, caraway, cardamom, chili powder, cinnamon, cloves, coriander, cumin, fennel, ginger, nutmeg, salt, star anise) of an Indian spice blend (garam masala) in cooked ground beef. Thiobarbituric acid (TBA) values were measured as an indicator of rancidity for cooked samples on 1-, 8-, or 15-d refrigerated storage. Cooked samples were evaluated by a trained panel (n= 13) for the intensity of rancid odor/flavor, beef flavor, and spice flavor and correlated with TBA values of same day samples. We also investigated possible additive effects between spice antioxidants and iron binding (type II) antioxidants on lipid oxidation by measuring TBA values. All spices had antioxidant effects on cooked ground beef, compared to controls. Among spices, cloves were the most effective in controlling lipid oxidation, with TBA values of 0.75, after 15-d refrigerated storage. All spices at their recommended levels lowered rancid odor and flavor in cooked ground beef, compared to controls. As expected, most spices also imparted distinctive flavors to cooked ground beef. There was a positive correlation (0.77) between TBA values on 15-d refrigerated storage and rancid odor/flavor. Type II antioxidants (iron-binding phosphate compounds) were more effective than individual type I antioxidants (spices and butylated hydroxytoluene; BHT) for maintenance of low TBA values in cooked ground beef during storage. Additive effects were observed with rosemary + milk mineral or sodium tripolyphosphate (STP) compared to rosemary alone. [source] Curcumin reduces the toxic effects of iron loading in rat liver epithelial cellsLIVER INTERNATIONAL, Issue 1 2009Donald J. Messner Abstract Background/Aims: Iron overload can cause liver toxicity and increase the risk of liver failure or hepatocellular carcinoma in humans. Curcumin (diferuloylmethane), a component of the food spice turmeric, has antioxidant, iron binding and hepatoprotective properties. The aim of this study was to quantify its effects on iron overload and the resulting downstream toxic effects in cultured T51B rat liver epithelial cells. Methods: T51B cells were loaded with ferric ammonium citrate (FAC) with or without the iron delivery agent 8-hydroxyquinoline. Cytotoxicity was measured by methylthiazolyldiphenyl-tetrazolium bromide assay. Iron uptake and iron bioavailability were documented by chemical assay, quench of calcein fluorescence and ferritin induction. Reactive oxygen species (ROS) were measured by a fluorescence assay using 2,,7,-dichlorodihydrofluorescein diacetate. Oxidative stress signalling to jnk, c-jun and p38 was measured by a Western blot with phospho-specific antibodies. Results: Curcumin bound iron, but did not block iron uptake or bioavailability in T51B cells given FAC. However, it reduced cytotoxicity, blocked the generation of ROS and eliminated signalling to cellular stress pathways caused by iron. Inhibition was observed over a wide range of FAC concentrations (50,500 ,M), with an apparent IC50 in all cases between 5 and 10 ,M curcumin. In contrast, desferoxamine blocked both iron uptake and toxic effects of iron at concentrations that depended on the FAC concentration. The effects of curcumin also differed from those of ,-tocopherol, which did not bind iron and was less effective at blocking iron-stimulated ROS generation. Conclusions: Curcumin reduced iron-dependent oxidative stress and iron toxicity in T51B cells without blocking iron uptake. [source] |