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Cristina Cereda Emanuela Leoni Pamela Milani Orietta Pansarasa Giuliano Mazzini Stefania Guareschi Elena Alvisi Andrea Ghiroldi Luca Diamanti Stefano Bernuzzi Mauro Ceroni Emanuela Cova 《PloS one》2013,8(10)
Several lines of evidence support the hypothesis of a toxic role played by wild type SOD1 (WT-SOD1) in the pathogenesis of sporadic amyotrophic lateral sclerosis (SALS). In this study we investigated both distribution and expression profile of WT-SOD1 in leukocytes from 19 SALS patients and 17 healthy individuals. Immunofluorescence experiments by confocal microscopy showed that SOD1 accumulates in the nuclear compartment in a group of SALS subjects. These results were also confirmed by western blot carried out on soluble nuclear and cytoplasmic fractions, with increased nuclear SOD1 level (p<0.05). In addition, we observed the presence of cytoplasmic SOD1 aggregates in agreement with an increased amount of the protein recovered by the insoluble fraction. A further confirmation of the overall increased level of SOD1 has been obtained from single cells analysis using flow cytometry as cells from SALS patients showed an higher SOD1 protein content (p<0.05). These findings add further evidence to the hypothesis of an altered WT-SOD1 expression profile in peripheral blood mononuclear cells (PBMCs) from patients with ALS suggesting that WT-SOD1 species with different degrees of solubility could be involved in the pathogenesis of the disease. 相似文献
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Controlled generation of reactive oxygen species orchestrates numerous physiological signaling events (Finkel, T. (2011) Signal transduction by reactive oxygen species. J. Cell Biol. 194, 7–15). A major cellular target of reactive oxygen species is the thiol side chain (RSH) of Cys, which may assume a wide range of oxidation states (i.e. −2 to +4). Within this context, Cys sulfenic (Cys-SOH) and sulfinic (Cys-SO2H) acids have emerged as important mechanisms for regulation of protein function. Although this area has been under investigation for over a decade, the scope and biological role of sulfenic/sulfinic acid modifications have been recently expanded with the introduction of new tools for monitoring cysteine oxidation in vitro and directly in cells. This minireview discusses selected recent examples of protein sulfenylation and sulfinylation from the literature, highlighting the role of these post-translational modifications in cell signaling. 相似文献
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Here we report massive seed predation of Pseudobombax grandiflorum (Bombacaceae) by Botogeris versicolurus (Psittacidae) in a forest fragment in Brazil. The intensity of seed predation was very high when compared to other studies in continuous forest, perhaps resulting from a scarcity of resources in such areas. This scarcity may limit the range of parrot's diet to a few plant species. It suggests that studies of Psittacidae seed predation may be important for conservation of some plants in fragments. 相似文献
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Summary The fermentation of grape must by Candida utilis ISS 28 was studied at different substrate concentrations, pH values, and nutrient supplementation in a shaken-flask fermenter, by using a composite design experiment.The experimental biomass yields were fitted to the only statistically significant factors with a mean standard error less than 8%, by using multiple regression analysis.Optimal conditions for maximum cell yield were established by plotting a series of loci at constant biomass yield and then verified experimentally, thus confirming the remarkable accuracy of the model 相似文献
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Mauro S. Sandrin K. Erin Lovering George Tachas Peter R. Collins Ian F. C. McKenzie 《Immunogenetics》1987,25(5):279-283
Human DNA was transfected into mouse L cells and tk+ HuLy-m2+ (= CD7+) transfectants isolated after growth in hypoxanthine, aminopterin, thymidine medium and repeated cloning. After several cycles of transfection, > 90% of HuLy-m2+ L cells could be detected, by rosetting and by cytofluorography, which showed the transfectants to have a density of CD7 two to five times that found on peripheral blood lymphocytes. Despite this, the 37 kd CD7+ dimer could only be identified with difficulty using cell-surface radioiodination and sodium dodecyl sulfate-polyacrylamide gel electrophoresis techniques. An antiserum was produced (C3H anti-HuLy-m2+ L cells) which, after absorption, was shown to react with HuLy-m2 antigens present on human thymocytes and lymphocytes and on CD7+ transfected L cells.Abbreviations BSA
bovine serum albumin
- DME
Dulbecco's modified Eagle's medium
- EDTA
ethylenediamine-tetraacetate
- HAT
hypoxanthine, aminopterin, thymidine
- HSV
herpes simplex virus
- PBL
peripheral blood lymphocyte
- PBS
phosphate-buffered saline
- RFC
rosette-forming cell
- SDS-PAGE
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
- tk
thymidine kinase 相似文献
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Heme pocket interactions in cytochrome c peroxidase studied by site-directed mutagenesis and resonance Raman spectroscopy 总被引:1,自引:0,他引:1
Resonance Raman spectra are reported for FeII and FeIII forms of cytochrome c peroxidase (CCP) mutants prepared by site-directed mutagenesis and cloning in Escherichia coli. These include the bacterial "wild type", CCP(MI), and mutations involving groups on the proximal (Asp-235----Asn, Trp-191----Phe) and distal (Trp-51----Phe, Arg-48----Leu and Lys) side of the heme. These spectra are used to assess the spin and ligation states of the heme, via the porphyrin marker band frequencies, especially v3, near 1500 cm-1, and, for the FeII forms, the status of the Fe-proximal histidine bond via its stretching frequency. The FeII-His frequency is elevated to approximately 240 cm-1 in CCP(MI) and in all of the distal mutants, due to hydrogen-bonding interactions between the proximal His-175 N delta and the carboxylate acceptor group on Asp-235. The FeII-His RR band has two components, at 233 and 246 cm-1, which are suggested to arise from populations having H-bonded and deprotonated imidazole; these can be viewed in terms of a double-well potential involving proton transfer coupled to protein conformation. The populations shift with changing pH, possibly reflecting structure changes associated with protonation of key histidine residues, and are influenced by the Leu-48 and Phe-191 mutations. A low-spin FeII form is seen at high pH for the Lys-48, Leu-48, Phe-191, and Phe-51 mutants; for the last three species, coordination of the distal His-52 is suggested by a approximately 200-cm-1 RR band assignable to Fe(imidazole)2 stretching.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献