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41.
Natalello A Prokorov VV Tagliavini F Morbin M Forloni G Beeg M Manzoni C Colombo L Gobbi M Salmona M Doglia SM 《Journal of molecular biology》2008,381(5):1349-1361
The existence of several prion strains and their capacity of overcoming species barriers seem to point to a high conformational adaptability of the prion protein. To investigate this structural plasticity, we studied here the aggregation pathways of the human prion peptide PrP82-146, a major component of the Gerstmann-Sträussler-Scheinker amyloid disease.By Fourier transform infrared (FT-IR) spectroscopy, electron microscopy, and atomic force microscopy (AFM), we monitored the time course of PrP82-146 fibril formation. After incubation at 37 °C, the unfolded peptide was found to aggregate into oligomers characterized by intermolecular β-sheet infrared bands. At a critical oligomer concentration, the emergence of a new FT-IR band allowed to detect fibril formation. A different intermolecular β-sheet interaction of the peptides in oligomers and in fibrils is, therefore, detected by FT-IR spectroscopy, which, in addition, suggests a parallel orientation of the cross β-sheet structures of PrP82-146 fibrils. By AFM, a wide distribution of PrP82-146 oligomer volumes—the smallest ones containing from 5 to 30 peptides—was observed. Interestingly, the statistical analysis of AFM data enabled us to detect a quantization in the oligomer height values differing by steps of ∼ 0.5 nm that could reflect an orientation of oligomer β-strands parallel with the sample surface. Different morphologies were also detected for fibrils that displayed high heterogeneity in their twisting periodicity and a complex hierarchical assembly.Thermal aggregation of PrP82-146 was also investigated by FT-IR spectroscopy, which indicated for these aggregates an intermolecular β-sheet interaction different from that observed for oligomers and fibrils. Unexpectedly, random aggregates, induced by solvent evaporation, were found to display a significant α-helical structure as well as several β-sheet components.All these results clearly point to a high plasticity of the PrP82-146 peptide, which was found to be capable of undergoing several aggregation pathways, with end products displaying different secondary structures and intermolecular interactions. 相似文献
42.
Beejan Asady Claudia F. Dick Karen Ehrenman Tejram Sahu Julia D. Romano Isabelle Coppens 《PLoS pathogens》2020,16(12)
Inorganic ions such as phosphate, are essential nutrients required for a broad spectrum of cellular functions and regulation. During infection, pathogens must obtain inorganic phosphate (Pi) from the host. Despite the essentiality of phosphate for all forms of life, how the intracellular parasite Toxoplasma gondii acquires Pi from the host cell is still unknown. In this study, we demonstrated that Toxoplasma actively internalizes exogenous Pi by exploiting a gradient of Na+ ions to drive Pi uptake across the plasma membrane. The Na+-dependent phosphate transport mechanism is electrogenic and functionally coupled to a cipargarmin sensitive Na+-H+-ATPase. Toxoplasma expresses one transmembrane Pi transporter harboring PHO4 binding domains that typify the PiT Family. This transporter named TgPiT, localizes to the plasma membrane, the inward buds of the endosomal organelles termed VAC, and many cytoplasmic vesicles. Upon Pi limitation in the medium, TgPiT is more abundant at the plasma membrane. We genetically ablated the PiT gene, and ΔTgPiT parasites are impaired in importing Pi and synthesizing polyphosphates. Interestingly, ΔTgPiT parasites accumulate 4-times more acidocalcisomes, storage organelles for phosphate molecules, as compared to parental parasites. In addition, these mutants have a reduced cell volume, enlarged VAC organelles, defects in calcium storage and a slightly alkaline pH. Overall, these mutants exhibit severe growth defects and have reduced acute virulence in mice. In survival mode, ΔTgPiT parasites upregulate several genes, including those encoding enzymes that cleave or transfer phosphate groups from phosphometabolites, transporters and ions exchangers localized to VAC or acidocalcisomes. Taken together, these findings point to a critical role of TgPiT for Pi supply for Toxoplasma and also for protection against osmotic stresses. 相似文献
43.
Concetta Fiore Javier Arrizon Anne Gschaedler Javier Flores Patrizia Romano 《World journal of microbiology & biotechnology》2005,21(6-7):1141-1147
Summary In Mexico there are different alcoholic beverages produced from agave juices from different agave plants, which are cooked,
fermented and distilled. For tequila production only Agave tequilana is allowed. In this study we compared yeast strains of different species from different origin (agave and grape juice) for
parameters of technological interest, such as SO2 and copper resistance, ethanol tolerance and enzymatic activities. All agave strains were found to be more resistant to SO2 and agave non-Saccharomyces yeasts were more tolerant to ethanol, whereas grape strains exhibited positive results for β-glucosidase and β-xylosidase activities. As regards fermentations of Agave tequilana juice with ethanol added at different concentrations, only agave Saccharomyces strains were more tolerant to ethanol than grape strains. 相似文献
44.
Biosynthesis of saturated and unsaturated fatty acids by a T-strain mycoplasma (Ureaplasma). 总被引:1,自引:3,他引:1 下载免费PDF全文
A human T mycoplasma (Ureaplasma urealyticum) incorporated radioactivity into its lipids from [1-14C]acetate in the growth medium. Methanolysis of the lipids showed the label to be confined almost entirely to the methyl esters of the fatty acids. About 80% of the label was associated with the methyl esters of the saturated fatty acids, and the rest was found in the unsaturated methyl ester fraction. Gas-liquid chromatography of the saturated methyl esters showed the label to be present in the peaks of palmitate, myristate, and stearate, whereas in the unsaturated methyl ester fraction most of the radioactivity emerged in the peak of palmitoleate. The addition of either oleic or palmitic acid to the growth medium markedly decreased the organisms' incorporation of radioactivity from acetate. It is concluded that the T mycoplasma strain is capable of de novo synthesis of both saturated and unsaturated fatty acids, in this respect differing from all of the Mycoplasma and Acholeplasma strains investigated to date. 相似文献
45.
Michela Castellani Raul Covian Thomas Kleinschroth Oliver Anderka Bernd Ludwig Bernard L. Trumpower 《The Journal of biological chemistry》2010,285(1):502-510
We previously proposed that the dimeric cytochrome bc1 complex exhibits half-of-the-sites reactivity for ubiquinol oxidation and rapid electron transfer between bc1 monomers (Covian, R., Kleinschroth, T., Ludwig, B., and Trumpower, B. L. (2007) J. Biol. Chem. 282, 22289–22297). Here, we demonstrate the previously proposed half-of-the-sites reactivity and intermonomeric electron transfer by characterizing the kinetics of ubiquinol oxidation in the dimeric bc1 complex from Paracoccus denitrificans that contains an inactivating Y147S mutation in one or both cytochrome b subunits. The enzyme with a Y147S mutation in one cytochrome b subunit was catalytically fully active, whereas the activity of the enzyme with a Y147S mutation in both cytochrome b subunits was only 10–16% of that of the enzyme with fully wild-type or heterodimeric cytochrome b subunits. Enzyme with one inactive cytochrome b subunit was also indistinguishable from the dimer with two wild-type cytochrome b subunits in rate and extent of reduction of cytochromes b and c1 by ubiquinol under pre-steady-state conditions in the presence of antimycin. However, the enzyme with only one mutated cytochrome b subunit did not show the stimulation in the steady-state rate that was observed in the wild-type dimeric enzyme at low concentrations of antimycin, confirming that the half-of-the-sites reactivity for ubiquinol oxidation can be regulated in the wild-type dimer by binding of inhibitor to one ubiquinone reduction site. 相似文献
46.
Vrabec R Vatavuk Z Pavlović D Sesar A Cala S Mandić K Bućan K 《Collegium antropologicum》2005,29(Z1):95-98
The aim of this paper was to evaluate the ocular findings in patients with chronic renal failure (CRF) undergoing haemodialysis (HD). In 64 patients undergoing haemodialysis (30 female and 34 male), aged 24-83 years (mean 58 years) on haemodialysis 1-213 months (mean 47 months) complete ocular examination were performed: visual acuity (VA), intraocular pressure (IOP), biomicroscopic examination and fundoscopy. On right eye sixty-nine percent of patents had VA 0.6 or better, and on left eye 84% of patients had VA 0.6 or better. Mean IOP before dialysis was 15 mmHg and after dialysis was 14 mmHg. In 9 patients (14%) we found corneo-conjunctival calcium deposits. No correlation of ocular calcification and parathyroid hormone (PTH) level or calcium and phosphate product were observed. 39 (60%) patients had cataract. Hypertensive vascular changes were seen in 44 (68%) patients and in 6 (7%) patients age-related macular degeneration. Seven patients had diabetes mellitus and in 5 diabetic retinopathy was observed. Patients with CRF or who are receiving HD represent unique group of patients. Pathologic change could be found in many tissue and organs, therefore we suggest ocular examination more frequently in dialysis patients. 相似文献
47.
The role of Mre11 phosphorylation in the cellular response to DNA double-strand breaks (DSBs) is not well understood. Here, we show that phosphorylation of Mre11 at SQ/TQ motifs by PIKKs (PI3 Kinase-related Kinases) induces MRN (Mre11–Rad50–Nbs1) complex dissociation from chromatin by reducing Mre11 affinity for DNA. Whereas phosphorylation of Mre11 at these residues is not required for DSB-induced ATM (Ataxia-Telangiectasia mutated) activation, abrogation of Mre11 dephosphorylation impairs ATM signaling. Our study provides a functional characterization of the DNA damage-induced Mre11 phosphorylation, and suggests that MRN inactivation participates in the down-regulation of damage signaling during checkpoint recovery following DSB repair. 相似文献
48.
Nicholas Romano Chaoshu Zeng 《Comparative biochemistry and physiology. Toxicology & pharmacology : CBP》2010,151(2):222-228
Ammonia-N toxicity to early Portunus pelagicus juveniles at different salinities was investigated along with changes to haemolymph osmolality, Na+, K+, Ca2+ and ammonia-N levels, ammonia-N excretion and gill Na+/K+-ATPase activity. Experimental crabs were acclimated to salinities 15, 30 and 45‰ for one week and 25 replicate crabs were subsequently exposed to 0, 20, 40, 60, 80, 100 and 120 mg L− 1 ammonia-N for 96-h, respectively. High ammonia-N concentrations were used to determine LC50 values while physiological measurements were conducted at lower concentrations. When crabs were exposed to ammonia-N, anterior gill Na+/K+-ATPase activity significantly increased (p < 0.05) at all salinities, while this only occurred on the posterior gills at 30‰. For crabs exposed to 20 and 40 mg L− 1 ammonia-N, both posterior gill Na+/K+-ATPase activity and ammonia-N excretion were significantly higher at 15‰ than those at 45‰. Despite this trend, the 96-h LC50 value at 15‰ (43.4 mg L− 1) was significantly lower (p < 0.05) than at both 30‰ and 45‰ (65.8 and 75.2 mg L− 1, respectively). This may be due to significantly higher (p < 0.05) haemolymph ammonia-N levels of crabs at low salinities and may similarly explain the general ammonia-N toxicity pattern to other crustacean species. 相似文献
49.
Crottini A Galimberti A Boto A Serra L Liu Y Yeung C Yang X Barbuto M Casiraghi M 《Molecular phylogenetics and evolution》2010,57(3):1312-1318
Paradoxornis webbianus and Paradoxornis alphonsianus naturally occur in South-East Asia. Due to a recent introduction, a mixed population currently occurs in northern Italy. A preliminary phylogeographic analysis using samples from Italy and China found little genetic differentiation between the two taxa and revealed the existence of two molecular lineages, sympatric in some part of China, that do not correspond to the morphological classification. Possible taxonomic changes and preliminary inferences on the relationships between Chinese and the Italian populations and on the likely provenance of the founders introduced in Italy are also discussed. 相似文献
50.
Pasquale V Romano VJ Rupnik M Dumontet S Cižnár I Aliberti F Mauri F Saggiomo V Krovacek K 《Folia microbiologica》2011,56(5):431-437
This pilot study was carried out to evaluate the occurrence of Clostridium difficile in marine environments and in edible shellfish. Samples of seawater, sediment, and zooplankton were collected at five sampling
stations in the Gulf of Naples. Six samples of edible shellfish, furthermore, were obtained: two from mussel farms and four
from wholesalers. The isolation and the characterization of C. difficile strains were carried out using selective media and molecular techniques, respectively. C. difficile was isolated from nine of the 21 samples investigated. Shellfish and zooplankton showed the highest prevalence of positive
samples. No C. difficile was detected in marine sediment. Majority of the C. difficile isolates were toxin A/B positive. Six known different PCR ribotypes (003, 005, 009, 010, 056, and 066) were identified, whereas
one strain may represent a new PCR ribotype. C. difficile may be present in the marine environment in Southern Italy, including shellfish and zooplankton. This study is reporting
the isolation of C. difficile from zooplankton, clams, and mussels and pointing out a new possible route to exposure to C. difficile of healthy individuals in the community. 相似文献