Sodium Nitroprusside (SNP) and S-Nitrosoglutathione (GSNO) differently affect mitochondrial H2O2 release at Complex-I. mM SNP increases while GSNO decreases the release induced by succinate alone or added on top of NAD-linked
substrates. Stimulation likely depends on Nitric Oxide (.NO) (released by SNP but not by GSNO) inhibiting cytochrome oxidase and mitochondrial respiration. Preincubations with SNP
or high GSNO (10 mM plus DTE to increases its .NO release) induces an inhibition of the succinate dependent H2O2 production consistent with a .NO dependent covalent modification. However maximal inhibition of the succinate dependent H2O2 release is obtained in the presence of low GSNO (20–100 μM), but not with SNP. This inhibition appears independent of .NO release since μM GSNO does not affect mitochondrial respiration, or the H2O2 detection systems and its effect is very rapid. Inhibition may be partly due to an increased removal of O2.− since GSNO chemically competes with NBT and cytochrome C in O2.− detection. 相似文献
Similar to σ-hole interactions, the π-hole interaction has attracted much attention in recent years. According to the positive electrostatic potentials above and below the surface of inorganic heterocyclic compounds S2N2 and three SN2P2 isomers (heterocyclic compounds 1–4), and the negative electrostatic potential outside the X atom of XH3 (X = N, P, As), S2N2/SN2P2?XH3 (X = N, P, As) complexes were constructed and optimized at the MP2/aug-cc-pVTZ level. The X atom of XH3 (X = N, P, As) is almost perpendicular to the ring of the heterocyclic compounds. The π-hole interaction energy becomes greater as the trend goes from 1?XH3 to 4?XH3. These π-hole interactions are weak and belong to “closed-shell” noncovalent interactions. According to the energy decomposition analysis, of the three attractive terms, the dispersion energy contributes more than the electrostatic energy. The polarization effect also plays an important role in the formation of π-hole complexes, with the contrasting phenomena of decreasing electronic density in the π-hole region and increasing electric density outside the X atom of XH3 (X = N, P, As).
Graphical abstract Computed density difference plots for the complexes 3?NH 3 (a 1), 3?PH 3 (b 1), 3?AsH 3 (c 1) and electron density shifts for the complexes 3?NH 3 (a 2), 3?PH 3 (b 2),3?AsH 3 (c 2) on the 0.001 a.u. contour
The structures and stabilities of eleven N13+ and N13– isomers have been investigated with second-order Møller–Plesset (MP2) and density functional theory (DFT) methods. Five N13+ isomers and six N13– isomers are all reasonable local minima on their potential energy hypersurfaces. The most stable N13+ cation is structure C-2 with C2v symmetry, which contains a pentazole ring and two N4 open chains. It is different from those of the N7+ and N9+ clusters, but similar to the N11+ cluster. Meanwhile, the most stable N13– structure A-2 is composed of a pentazole ring and a six-membered ring connected by two nitrogen atoms. It is not only different from those of the N7– and N9– clusters, but also from the N11– cluster. The decomposition pathways of structures C-2 and A-2 were investigated at the B3LYP/(aug)-cc-pVDZ level. From the barrier heights of the structures C-2 and A-2 decomposition processes, it is suggested that C-2 is difficult to observe experimentally and A-2 may be observed as a short-lived species.
Figure Optimized geometrical parameters of N13+ isomer C-2相似文献
THE urate-binding α1–α2 globulin has been isolated from human plasma in a highly purified state1. The protein was purified by DEAE-‘Sephadex’, ammonium sulphate precipitation and semi-preparative Polyacrylamide gel electrophoresis. The urate-binding α1–α2 globulin is a rod-shaped glycoprotein, containing 12.1% carbohydrate, with an isoelectric point of 4.6 and a molecular weight of 67,000 ± 4,000. Amino-acid analysis indicated an unknown basic compound which appeared as an extra peak just in front of lysine1. To identify this compound, high voltage paper electrophoresis has been carried out on a plate electrophoresis apparatus in pyridine-acetate buffer pH 3.5. A spot separated out corresponding to ornithine. Amino-acid analysis on a BC-200 automatic analyser (Bio-Cal Instruments Co., West Germany), with a 54 cm column at 55° C and with 0.35 M sodium citrate buffer, pH 5.28, as elution buffer at a flow-rate of 150 ml./h, showed that ornithine was present. The presence of ornithine in the protein hydrolysate was also verified by gas chromatography/mass spectrometry2. 相似文献
While ~30% of the human genome encodes membrane proteins, only a handful of structures of membrane proteins have been resolved to high resolution. Here, we studied the structure of a member of the Cys-loop ligand gated ion channel protein superfamily of receptors, human type A γ2α1β2α1β2 gamma amino butyric acid receptor complex in a lipid bilayer environment. Studying the correlation between the structure and function of the gamma amino butyric acid receptor may enhance our understanding of the molecular basis of ion channel dysfunctions linked with epilepsy, ataxia, migraine, schizophrenia and other neurodegenerative diseases. The structure of human γ2α1β2α1β2 has been modeled based on the X-ray structure of the Caenorhabditis elegans glutamate-gated chloride channel via homology modeling. The template provided the first inhibitory channel structure for the Cys-loop superfamily of ligand-gated ion channels. The only available template structure before this glutamate-gated chloride channel was a cation selective channel which had very low sequence identity with gamma aminobutyric acid receptor. Here, our aim was to study the effect of structural corrections originating from modeling on a more reliable template structure. The homology model was analyzed for structural properties via a 100 ns molecular dynamics (MD) study. Due to the structural shifts and the removal of an open channel potentiator molecule, ivermectin, from the template structure, helical packing changes were observed in the transmembrane segment. Namely removal of ivermectin molecule caused a closure around the Leu 9 position along the ion channel. In terms of the structural shifts, there are three potential disulfide bridges between the M1 and M3 helices of the γ2 and 2 α1 subunits in the model. The effect of these disulfide bridges was investigated via monitoring the differences in root mean square fluctuations (RMSF) of individual amino acids and principal component analysis of the MD trajectory of the two homology models—one with the disulfide bridge and one with protonated Cys residues. In all subunit types, RMSF of the transmembrane domain helices are reduced in the presence of disulfide bridges. Additionally, loop A, loop F and loop C fluctuations were affected in the extracellular domain. In cross-correlation analysis of the trajectory, the two model structures displayed different coupling in between the M2–M3 linker region, protruding from the membrane, and the β1-β2/D loop and cys-loop regions in the extracellular domain. Correlations of the C loop, which collapses directly over the bound ligand molecule, were also affected by differences in the packing of transmembrane helices. Finally, more localized correlations were observed in the transmembrane helices when disulfide bridges were present in the model. The differences observed in this study suggest that dynamic coupling at the interface of extracellular and ion channel domains differs from the coupling introduced by disulfide bridges in the transmembrane region. We hope that this hypothesis will be tested experimentally in the near future. 相似文献
Utilizing first-principles calculations, we studied the electronic and optical properties of C24, C12X6Y6, and X12Y12 fullerenes (X?=?B, Al; Y?=?N, P). These fullerenes are energetically stable, as demonstrated by their negative cohesive energies. The energy gap of C24 may be tuned by doping, and the B12N12 fullerene was found to have the largest energy gap. All of the fullerenes had finite optical gaps, suggesting that they are optical semiconductors, and they strongly absorb UV radiation, so they could be used in UV light protection devices. They could also be used in solar cells and LEDs due to their low reflectivities.
Mitochondrial production of H2O2 is low with NAD substrates (glutamate/pyruvate, 3 and 2 mM) (G/P) and increases over ten times upon further addition of succinate,
with the formation of a sigmoidal curve (semimaximal value at 290 μM, maximal H2O2 production at 600 μM succinate). Malate counteracts rapidly the succinate induced increased H2O2 release and moves the succinate dependent H2O2 production curve to the right. Nitric oxide (NO) and carbon monoxide (CO) are cytochrome c oxidase inhibitors which increase
mitochondrial ROS production. Cyanide (CN−) was used to mimic NO and CO. In the presence of G/P and succinate (300 μM), CN− progressively increased the H2O2 release rate, starting at 1.5 μM. The succinate dependent H2O2 production curve was moved to the left by 30 μM CN−. The Vmax was little modified. We conclude that succinate is the controller of mitochondrial H2O2 production, modulated by malate and CN−. We propose that succinate promotes an interaction between Complex II and Complex I, which activates O2− production. 相似文献
Nine minima were found on the intermolecular potential energy surface for the ternary system HNO3(CH3OH)2 at the MP2/aug-cc-pVDZ level of theory. The cooperative effect, which is a measure of the hydrogen-bonding strength, was probed in these nine conformations of HNO3…(CH3OH)2. The results are discussed here in terms of structures, energetics, infrared vibrational frequencies, and topological parameters. The cooperative effect was observed to be an important contributor to the total interaction energies of the cyclic conformers of HNO3…(CH3OH)2, meaning that it cannot be neglected in simulations in which the pair-additive potential is applied.
PROSTAGLANDIN (PG) F2αhas antifertility effects in many species1–3 but there are conflicting suggestions as to its mechanism of action. For example, it may cause the degeneration of the corpus luteum by decreasing blood flow in the uteroovarian vein4; alternatively, its action may be due to a hypersecretion of luteinizing hormone (LH) by the pituitary3,5. I have investigated the effects of PGF2α, E2 and E1 on pregnancy in mice and examined the mechanism of action of PGF2α. 相似文献
D2− ions produced in collisions of D− ions with relative energies of 2.5–9.2 eV were detected for the first time. It is shown that the effective cross section
for this reaction is no less than 1.5 × 10−14 cm2. Along with the theoretically predicted short-lived state of negative molecular deuterium ions, a state existing for more
than 1 μs was observed. 相似文献
Ab initio calculations have been performed using the complete basis set model (CBS-QB3) to study the reaction mechanism of
butane radical (C4H9•) with oxygen (O2). On the calculated potential energy surface, the addition of O2 to C4H9• forms three intermediates barrierlessly, which can undergo subsequent isomerization or decomposition reaction leading to
various products: HOO• + C4H8, C2H5• + CH2CHOOH, OH• + C3H7CHO, OH• + cycle-C4H8O, CH3• + CH3CHCHOOH, CH2OOH• + C3H6. Five pathways are supposed in this study. After taking into account the reaction barrier and enthalpy, the most possible
reaction pathway is C4H9• + O2 → IM1 → TS5 → IM3 → TS6 → IM4 → TS7 → OH• + cycle-C4H8O. 相似文献
A preliminary study on the interaction of G protein (guanine triphosphate binding pro- tein) β1γ2 subunits and their coupled components in cell signal transduction was conducted in vitro. The insect cell lines, Sf9 (Spodoptera frugiperda) and H5 (Trichoplusia ni) were used to express the recombinant protein Gβ1γ2. The cell membrane containing Gβ1γ2 was isolated through affinity chromatography column with Ni-NTA agarose by FPLC method, and the highly purified protein was obtained. The adenylyl cyclase 2 (AC2) activity assay showed that the purified Gβ1γ2 could significantly stimulate AC2 activity. The interaction of β1γ2 subunits of G protein with the cytoplasmic tail of various mammalian adenylyl cyclases was monitored by BIAcore technology using NTA sensor chip, which relies on the phenomenon of surface plasmon resonance (SPR). The experiments showed the direct binding of Gβ1γ2 to the cytoplasmic tail C2 domain of AC2. The specific binding domain of AC2 with Gβ1γ2 was the same as AC2 activity domain which was stimulated by β1γ2. 相似文献
To analyze the influence of the beta-subunit on the kinetic properties of GlyR channel currents, alpha(1)-subunits and alpha(1)beta-subunits were transiently expressed in HEK 293 cells. A piezo dimorph was used for fast application of glycine to outside-out patches. The rise time of activation was dose dependent for both receptors and decreased with increasing glycine concentrations. Subunit composition had no effect on the time course of activation. Coexpression of alpha(1)- and beta-subunits resulted in a significantly lower EC(50) and a reduced slope of the dose-response curve of glycine compared with expression of alpha(1)-subunits alone. For both receptor subtypes, the time course of desensitization was concentration dependent. Desensitization was best fitted with a single time constant at 10-30 micro M, with two at 0.1 mM, and at saturating concentrations (0.3-3 mM) with three time constants. Desensitization of homomeric alpha(1)-receptor channels was significantly slower than that of alpha(1)beta-receptor channels. The time course of current decay after the end of glycine pulses was tested at different pulse durations of 1 mM glycine. It was best fitted with two time constants for both alpha(1) and alpha(1)beta GlyR channels, and increased significantly with increasing pulse duration. 相似文献
In two mountain ecosystems at the Alptal research site in central Switzerland, pulses of 15NO3 and 15NH4 were separately applied to trace deposited inorganic N. One forested and one litter meadow catchment, each approximately
1600 m2, were delimited by trenches in the Gleysols. K15NO3 was applied weekly or fortnightly over one year with a backpack sprayer, thus labelling the atmospheric nitrate deposition.
After the sampling and a one-year break, 15NH4Cl was applied as a second one-year pulse, followed by a second sampling campaign. Trees (needles, branches and bole wood),
ground vegetation, litter layer and soil (LF, A and B horizon) were sampled at the end of each labelling period. Extractable
inorganic N, microbial N, and immobilised soil N were analysed in the LF and A horizons. During the whole labelling period,
the runoff water was sampled as well. Most of the added tracer remained in both ecosystems. More NO3− than NH4+ tracer was retained, especially in the forest. The highest recovery was in the soil, mainly in the organic horizon, and in
the ground vegetation, especially in the mosses. Event-based runoff analyses showed an immediate response of 15NO3− in runoff, with sharp 15N peaks corresponding to discharge peaks. NO3− leaching showed a clear seasonal pattern, being highest in spring during snowmelt. The high capacity of N retention in these
ecosystems leads to the assumption that deposited N accumulates in the soil organic matter, causing a progressive decline
of its C:N ratio. 相似文献
The nucleation, ice crystal shapes and thermodynamic stability of polar stratospheric clouds particles are interesting concerns owing to their implication in the ozone layer destruction. Some of these particles are formed by conformers of H2O, HNO3, and H2SO4. We carried out calculations using density functional theory (DFT) to obtain optimized structures. Several stable trimers are achieved —divided in two groups, one with HNO3 moiety, second with H2SO4 moiety— after pre-optimization at B3LYP/6-31G and subsequently optimization at B3LYP/aug-cc-pVTZ level of theory. For both most stable conformers five H2O molecules are added to their optimized trimers to calculate hydrated geometries. The OH stretching harmonic frequencies are provided for all aggregates. The zero-point energy correction (ZEPC), relative electronic energies (?E), relative reaction Gibbs free energies ?(?G)k-relative, and cooling constant (Kcooling) are reported at three temperatures: 188 K, 195 K, and 210 K. Shapes given in our calculations are compared with various experimental shapes as well as comparisons with their thermo-stabilities.
Graphical Abstract Facet shapes and thermo-stabilities of H2SO4?HNO3 hydrates involved in polar stratospheric clouds. IR spectrum of WNS-1+5W structure and its circular facet
The ability of an ecosystem to retain anthropogenic nitrogen (N) deposition is dependent upon plant and soil sinks for N,
the strengths of which may be altered by chronic atmospheric N deposition. Sugar maple (Acer saccharum Marsh.), the dominant overstory tree in northern hardwood forests of the Lake States region, has a limited capacity to take
up and assimilate NO3−. However, it is uncertain whether long-term exposure to NO3− deposition might induce NO3− uptake by this ecologically important overstory tree. Here, we investigate whether 10 years of experimental NO3− deposition (30 kg N ha−1 y−1) could induce NO3− uptake and assimilation in overstory sugar maple (approximately 90 years old), which would enable this species to function
as a direct sink for atmospheric NO3− deposition. Kinetic parameters for NH4+ and NO3− uptake in fine roots, as well as leaf and root NO3− reductase activity, were measured under conditions of ambient and experimental NO3− deposition in four sugar maple-dominated stands spanning the geographic distribution of northern hardwood forests in the
Upper Lake States. Chronic NO3− deposition did not alter the Vmax or Km for NO3− and NH4+ uptake nor did it influence NO3− reductase activity in leaves and fine roots. Moreover, the mean Vmax for NH4+ uptake (5.15 μmol 15N g−1 h−1) was eight times greater than the Vmax for NO3− uptake (0.63 μmol 15N g−1 h−1), indicating a much greater physiological capacity for NH4+ uptake in this species. Additionally, NO3− reductase activity was lower than most values for woody plants previously reported in the literature, further indicating
a low physiological potential for NO3− assimilation in sugar maple. Our results demonstrate that chronic NO3− deposition has not induced the physiological capacity for NO3− uptake and assimilation by sugar maple, making this dominant species an unlikely direct sink for anthropogenic NO3− deposition. 相似文献
Summary Phosphatidylinositol 3-kinase (PI3K) pathway is important for platelet activation. Recent studies showed that PI3K and oscillative
calcium could cross talk to each other and positively regulate integrin α IIbβ3-mediated outside-in signaling. However, the mechanism of this feedback regulation remains to be further characterized. Here
we found that treatments of both PI3K inhibitor wortmannin and P2Y1 inhibitor A3P5P could inhibit granular secretion in platelets.
Additionally, when RGD-substrate adherent platelets were treated with the ADP scavenger apyrase to deplete the granular-released
ADP, their attachments in engaging with substrates became looser and the frequency of calcium oscillation decreased. Since
it is known that ADP stimulates the PI3K and calcium signal primarily through P2Y12 and P2Y1 receptors respectively, our data
indicated that integrin αIIbβ3 downstream PI3K and calcium activation might be not completely coupled to integrin associated signaling complex, but in part
through feedback stimulation by granular released ADP. Our data indicates the important roles of PI3K and granular released
ADP in coordinating the feedback regulations in integrin αIIbβ3-mediated platelet activation. 相似文献
A new quartz crystal microbalance immunoassay for the detection of carcinoembryonic antigen (CEA) was developed by means of
immobilizing anti-CEA onto magnetic CoFe2O4/SiO2 composite nanoparticles-functionalized biomimetic interface. Under optimal conditions, the frequency shift was proportional
to the CEA concentration in the range of 2.5–55 ng/mL with a detection limit of 0.5 ng/mL at a signal-to-noise ratio of 3.
Moreover, the immunosensor system showed an acceptable reproducibility and stability. Clinical serum specimens were assayed
with this method, and the results were in acceptable agreement with those obtained from ELISA. Compared with the conventional
ELISA assay, the proposed immunoassay system was simple and rapid without multiple labeling and separation steps. Importantly,
the developed immunoassay protocol could be further extended for the determination of other antigens. 相似文献
F0F1ATPsynthase is now known to be expressed as a plasma membrane receptor for several extracellular ligands. On hepatocytes,
ecto–F0F1ATPsynthase binds apoA–I and triggers HDL endocytosis concomitant with ATP hydrolysis. Considering that inhibitor protein
IF1 was shown to regulate the hydrolytic activity of ecto–F0F1ATPsynthase and to interact with calmodulin (CaM) in vitro, we investigated the subcellular distributions of IF1, calmodulin (CaM), OSCP and β subunits of F0F1ATPsynthase in HepG2 cells. Using immunofluorescence and Western blotting, we found that around 50% of total cellular IF1 is localized outside mitochondria, a relevant amount of which is associated to the plasma membrane where we also found Ca2+–CaM, OSCP and β. Confocal microscopy showed that IF1 colocalized with Ca2+–CaM on plasma membrane but not in mitochondria, suggesting that Ca2+–CaM may modulate the cell surface availability of IF1 and thus its ability to inhibit ATP hydrolysis by ecto–F0F1ATPsynthase. These observations support a hypothesis that the IF1–Ca2+–CaM complex, forming on plasma membrane, functions in the cellular regulation of HDL endocytosis by hepatocytes. 相似文献
Tumorigenic cell lines are more susceptible to [Re6Se8I6]3? cluster-induced death than normal cells, becoming a novel candidate for cancer treatment. Still, the feasibility of using this type of molecules in human patients remains unclear and further pharmacokinetics analysis is needed. Using coupled plasma optical emission spectroscopy, we determined the Re-cluster tissue content in injected mice, as a biodistribution measurement. Our results show that the Re-cluster successfully reaches different tissues, accumulating mainly in heart and liver. In order to dissect the mechanism underlying cluster biodistribution, we used three different experimental approaches. First, we evaluate the degree of lipophilicity by determining the octanol/water partition coefficient. The cluster mostly remained in the octanol fraction, with a coefficient of 1.86?±?0.02, which indicates it could potentially cross cell membranes. Then, we measured the biological membrane penetration through a parallel artificial membrane permeability assays (PAMPA) assay. The Re-cluster crosses the artificial membrane, with a coefficient of 122 nm/s that is considered highly permeable. To evaluate a potential application of the Re-cluster in central nervous system (CNS) tumors, we analyzed the cluster’s brain penetration by exposing cultured blood–brain-barrier (BBB) cells to increasing concentrations of the cluster. The Re-cluster effectively penetrates the BBB, reaching nearly 30% of the brain side after 24 h. Thus, our results indicate that the Re-cluster penetrates biological membranes reaching different target organs—most probably due to its lipophilic properties—becoming a promising anti-cancer drug with high potential for CNS cancer’s diagnosis and treatment. 相似文献