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931.
Uptake kinetics of iron-phytosiderophores in two maize genotypes differing in iron efficiency 总被引:5,自引:0,他引:5
Iron inefficiency in the maize ( Zea mays L.) mutant ysl is caused by a defect in the uptake system for Fe-phytosiderophores. To characterize this defect further, the uptake kinetics of Fe-phytosiderophores in ysl was compared to the Fe-efficient maize cultivar Alice. Short-term uptake of 59 Fe-labeled Fe-deoxymugineic acid (Fe-DMA) was measured over a concentration range of 0.03 to 300 μM. Iron uptake in Fe-deficient plants followed Michaelis-Menten kinetics up to about 30 μM and was linear at higher concentrations, indicating two kinetically distinct components in the uptake of Fe-phytosiderophores. The saturable component had similar Km (∼ 10 μM) in both genotypes. In contrast. Vmax was 5.5 μmol Fe-DMA g−1 dry weight [30 min]−1 in Alice, but only 0.6 μmol Fe-DMA g−1 dry weight [30 min]−1 in ysl . Uptake experiments with double-labeled 59 Fe-[14 C]DMA suggest that in both cultivars Fe-DMA was taken up by the roots as the intact chelate. The results indicate the existence of a high-affinity and a low-affinity uptake system mediating Fe-phytosiderophore transport across the root plasma membrane in maize. Apparently, the mutation responsible for Fe inefficiency in ysl affected high-affected uptake and led to a decrease in activity and/or number of Fe-phytosiderophore transporters. 相似文献
932.
Jeffrey M. Feldman Russell Hilf 《Biochimica et Biophysica Acta (BBA)/Molecular Cell Research》1985,845(2):265-271
Purified plasma membrane vesicles isolated from R3230AC rat mammary tumors displayed carrier-mediated and stereospecific uptake. Uptake was shown to be proportional to protein concentration, sensitive to increasing osmolarity, and inhibited only by substrates entering by the same carrier. Carrier-mediated glucose uptake was inhibited rapidly by estradiol-17β and phloretin in a dose-dependent manner, whereas proline uptake was not affected by estradiol-17β. The data suggest that the inhibition of glucose by estradiol and phloretin, originally observed in whole cells, occurs by an interaction of the steroid with a component on the plasma membrane. In contrast, the lack of effects of estradiol on proline transport into vesicles implies that intracellular components may have mediated the estrogen-induced effects observed in whole cells. 相似文献
933.
Five different strains of wine yeasts were investigated with respect to active uptake of [35S] sulfate and its regulation by methionine. Considerable differences exist between low and high sulfite-producing strains in the initial velocity of sulfate uptake. Further differences were established in repression of sulfate permease by l-methionine, most evident in a total lack of repression in one of the high sulfite producers. These findings explain in part variable sulfite and sulfide formation.List of Abbreviations CCMP
carbonylcyanide-p-trifluormethoxyphenylhydrazone 相似文献
934.
Andrei V. Astashkin Arnold M. Raitsimring F. Ann. Walker Christopher Rensing Megan M. McEvoy 《Journal of biological inorganic chemistry》2005,10(3):221-230
Electron paramagnetic resonance (EPR) spectroscopy has been used to structurally characterize the copper-binding site in CusF protein from Escherichia coli. The EPR spectra indicate a single type II copper center with parameters typical for nitrogen and oxygen ligands (A~200 G, g~2.186, g~2.051). The pulsed EPR data show that one of the ligands to Cu2+ is an imidazole ring of a histidine residue. The remote amino nitrogen of this imidazole ring is readily observed by electron spin-echo envelope modulation spectroscopy, while the imino nitrogen that is directly coordinated to the Cu2+ ion is observed by pulsed electron–nuclear double resonance (ENDOR). In addition, the ENDOR spectra reveal the presence of one more nitrogen ligand that was assigned to be a deprotonated peptide nitrogen. Apart from the two nitrogen ligands, it has been established that there are two nearby hydroxyl protons, although whether these belong to a single equatorial water ligand or two equatorial hydroxide ligands is not known.
相似文献
Megan M. McEvoyEmail: Phone: +1-520-6213489Fax: +1-520-6211697 |
935.
Further study has been made of metal-catalyzed oxidation (MCO) reactions and mass spectrometry as a method to determine the binding site of copper in metalloproteins. The role of ascorbate and a variety of oxidizing agents, including O2, H2O2, and S2O8(2-), have been investigated using Cu/Zn superoxide dismutase (SOD) as a model system. Ascorbate is found to play two competing roles in the MCO reactions. It reduces Cu(II), which initiates and maintains the generation of reactive oxygen species, and it scavenges radicals, which helps to localize oxidation products to amino acids near the metal center. An ascorbate concentration of 100 mM is found to be optimal with regard to localizing oxidation products to only the Cu-binding residues (His44, His46, His61, and His118) of Cu/Zn SOD. This concentration of ascorbate is very similar to the optimum concentration found in our previous studies of different Cu-binding proteins. Another notable result from this study is the observation that S2O8(2-) is more effective as an oxidant than O2 or H2O2 in the MCO reactions. Because S2O8(2-) is more stable in solution than H2O2, using it as an oxidizing agent results in much less nonspecific oxidation to the protein. The overall results of this study suggest that general MCO reaction conditions may exist for determining the metal-binding site of a wide range of Cu-binding proteins. 相似文献
936.
937.
Tetranuclear Cu(II) complexes of N-(2-hydroxymethylphenyl)salicylideneimine (H2L1-H) and its homologues (5-CH3: H2L1-Me, 5-Cl: H2L1-Cl), [Cu(L1-H)]4 · 3H2O (1), [Cu(L1-Me)]4 · 2CH2Cl2 (2), and [Cu(L1-Cl)]4 · 2CH2Cl2 (3), have been characterized by X-ray crystal structure analyses and magnetic measurements. The structure analyses revealed that the complexes 1-3 have a defective double-cubane tetra copper(II) core connected by μ3-alkoxo bridges. The intramolecular Cu?Cu distances are in the range from 5.251(2)-5.256(3) Å for the longest to 3.0518(9)-3.092(2) Å for the shortest. Each Cu(II) ion has a square-pyramidal geometry and the dihedral angles between adjacent Cu(II) basal planes are almost right angles. Magnetic measurements of the present complexes indicate that weak antiferromagnetic interactions (J=−15 to −19 cm−1) between neighboring copper(II) ions are dominant in these tetracopper cores. 相似文献
938.
Calcium Enhances In Vitro Free Radical-Induced Damage to Brain Synaptosomes, Mitochondria, and Cultured Spinal Cord Neurons 总被引:1,自引:1,他引:1
Preincubation of rat brain synaptosomes with xanthine and xanthine oxidase (X/XO) in Ca2+-free Krebs buffer resulted in a 27% inhibition of synaptosomal gamma-aminobutyric acid (GABA) uptake. Addition of 1.5 mM CaCl2 increased the inhibition with X/XO to 46%, and inhibition was essentially complete when the calcium ionophore A23187 also was included. In other studies, preincubation of purified rat brain mitochondria with the combination of X/XO and 4 microM CaCl2 produced a significant (38%) decrease in state 3 respiration with glutamate/malate as substrate that was not seen with either X/XO or Ca2+ alone. Similar results were obtained using cultured mouse spinal cord neurons in which incubation with X/XO/ADP/FeCl2 and A23187 produced membrane damage as assessed by a 32% reduction of neuronal Na+, K+-ATPase activity. Neither X/XO/ADP/FeCl2 nor A23187 alone caused detectable inhibition. These results demonstrate the synergistic damaging effect of free radicals and Ca2+ on membrane function. In addition, they suggest that free radical-induced peroxidation of membrane lipid, occurring focally during complete or nearly complete ischemia in vivo, could result in intense cellular perturbation when coupled with increased intracellular Ca2+. 相似文献
939.
Pseudomonas putida IFO13696, a recombinant strain with dsz desulfurization genes, desulfurized dibenzothiophene (DBT) in water but not in n-tetradecane. By introducing into this recombinant strain the hcuABC genes that take part in the uptake of DBT in the oil phase into the cell, 82% of 1 mM DBT in n-tetradecane was degraded in 24 h by resting cells. The products of hcuABC genes thus acted in the uptake of DBT in n-tetradecane into the cells and were effective in desulfurization of DBT in the hydrocarbon phase. 相似文献
940.
Following a toxic flood from a mine spill that affected over 45 km2 in Southern Spain, experiments were conducted in 1999 to test the feasibility of using crops for phytoremediation of the area, after the mechanical removal of the mud. Two cereals, barley and triticale, and two Brassicaspp., rapeseed and ethiopian mustard, were planted in three contaminated plots, 50 × 100 m each, and in a control plot outside the affected area. Soil and plant contents of As, Cd, Cu, Pb, Tl and Zn were measured and bioaccumulation coefficients (BC) were calculated at maturity. The four crops tested accumulated Cd and Zn in the above-ground biomass only in the plot on acid soil. Both species of Brassica accumulated Tl (average BC of 3.6 and 1.4 for rapeseed and mustard, respectively, in contaminated plots). None of the four crop plants accumulated As, Cu and Pb under the experimental conditions. Maximum plant uptake values from soil were 5.4 mg m–2 of As, 0.54 mg m–2 of Cd, 9.7 mg m–2 of Cu, 7.0 mg m–2 of Pb, 3.4 mg m–2 of Tl, and 260 mg m–2 of Zn. Total crop uptake gave estimates for successful phytoremediation of at least five decades, casting doubts on the feasibility of using these crops for decontamination of the area. Nevertheless, cereal grains had mineral contents below toxicity levels for livestock, therefore it might be possible to use these crops for livestock feed while reducing deep percolation and gradually removing metals from polluted soils. 相似文献