The interaction of the diastereomeric complexes Λ-[Ru(bpy)2(m-GHK)]Cl2 and Δ-[Ru(bpy)2(m-GHK)]Cl2 (bpy is 2,2′-bipyridine, GHK is glycine–l-histidine–l-lysine) with the deoxynucleotide duplex d(5′-CGCGATCGCG)2 was studied by means of 1H NMR spectroscopy. At a Δ-isomer to DNA ratio of 1:1, significant shifts for the metal complex are observed, whereas there is negligible effect on
the oligonucleotide protons and only one intermolecular nuclear Overhauser effect (NOE) is present at the 2D nuclear Overhauser
enhancement spectroscopy spectrum. The 1Η NMR spectrum at ratio 2:1 is characterized by a slight shift for the Δ-isomer’s bpy aromatic protons as well as significant shifts for the decanucleotide G4 H1′ and Η2″, A5 H2, G10 H1′, T6 NH and G2 NH protons. Furthermore, at ratio 2:1, 11 intermolecular NOEs are observed. The majority of the NOEs involve the sugar Η2′
and Η2″ protons sited in the major groove of the decanucleotide. Increasing the Δ-isomer to d(CGCGATCGCG)2 ratio to 5:1 results in noteworthy spectral changes. The Δ-isomer’s proton shifts are reduced, whereas significant shifts are observed for the decanucleotide protons, especially the
sugar protons, as well as for the exchangeable protons. Interaction is characterized by the presence of only one intermolecular
NOE. Furthermore, there is significant broadening of the imino proton signals as the ratio of the Δ-isomer to DΝΑ increases, which is attributed to the opening of the two strands of the duplex. The Λ-isomer, on the other hand, approaches the minor groove of the oligonucleotide and interacts only weakly, possibly by electrostatic
interactions. Photocleavage studies were also conducted with the plasmid pUC19 and a 158-bp restriction fragment, showing
that both diastereomers cleave DNA with similar efficiency, attacking mainly the guanines of the sequence probably by generating
active oxygen species.
Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users.
An erratum to this article can be found at 相似文献
The nature of M–M bonding and aromaticity of [M2(NHCHNH)3]2(μ-E)2 (E?=?O, S; M?=?Nb, Mo, Tc, Ru, Rh) was investigated using atoms in molecules (AIM) theory, electron localization function (ELF), natural bond orbital (NBO) and molecular orbital analysis. These analyses led to the following main conclusions: in [M2(NHCHNH)3]2(μ-E)2 (E?=?O, S; M?=?Nb, Mo, Tc, Ru, Rh), the Nb–Nb, Ru–Ru, and Rh–Rh bonds belong to “metallic” bonds, whereas Mo–Mo and Tc–Tc drifted toward the “dative” side; all these bonds are partially covalent in character. The Nb–Nb, Mo–Mo, and Tc–Tc bonds are stronger than Ru–Ru and Rh–Rh bonds. The M–M bonds in [M2(NHCHNH)3]2(μ-S)2 are stronger than those in [M2(NHCHNH)3]2(μ-O)2 for M?=?Nb, Mo, Tc, and Ru. The NICS(1)ZZ values show that all of the studied molecules, except [Ru2(NHCHNH)3]2(μ-O)2, are aromaticity molecules. O-bridged compounds have more aromaticity than S-bridged compounds.
Graphical AbstractLeft Molecular graph, and right electron localization function (ELF) isosurface of [M2(NHCHNH)3]2(μ-E)2(E?=?O, S; M?=?Nb, Mo, Tc, Ru, Rh)
We present two NMR experiments, (3,2)D HNHA and (3,2)D HNHB, for rapid and accurate measurement of 3J(H N-H alpha) and 3J(N-H beta) coupling constants in polypeptides based on the principle of G-matrix Fourier transform NMR spectroscopy and quantitative J-correlation. These experiments, which facilitate fast acquisition of three-dimensional data with high spectral/digital resolution and chemical shift dispersion, will provide renewed opportunities to utilize them for sequence specific resonance assignments, estimation/characterization of secondary structure with/without prior knowledge of resonance assignments, stereospecific assignment of prochiral groups and 3D structure determination, refinement and validation. Taken together, these experiments have a wide range of applications from structural genomics projects to studying structure and folding in polypeptides. 相似文献
We present a highly sensitive pulse sequence, carbonyl carbon label selective 1H–15N HSQC (CCLS-HSQC) for the detection of signals from 1H–15N units involved in 13C′–15N linkages. The CCLS-HSQC pulse sequence utilizes a modified 15N CT evolution period equal to 1/(
) (∼33 ms) to select for 13C′–15N pairs. By collecting CCLS-HSQC and HNCO data for two proteins (8 kDa ubiquitin and 20 kDa HscB) at various temperatures
(5–40°C) in order to vary correlation times, we demonstrate the superiority of the CCLS-HSQC pulse sequence for proteins with
long correlation times (i.e. higher molecular weight). We then show that the CCLS-HSQC experiment yields assignments in the
case of a 41 kDa protein incorporating pairs of 15N- and 13C′-labeled amino acids, where a TROSY 2D-HN(CO) had failed. Although the approach requires that the 1H–15N HSQC cross peaks be observable, it does not require deuteration of the protein. The method is suitable for larger proteins
and is less affected by conformational exchange than HNCO experiments, which require a longer period of transverse 15N magnetization. The method also is tolerant to the partial loss of signal from isotopic dilution (scrambling). This approach
will be applicable to families of proteins that have been resistant to NMR structural and dynamic analysis, such as large
enzymes, and partially folded or unfolded proteins. 相似文献
The frequency of individuals with empty stomachs (FES) can vary greatly among northern pike populations. However, the FES
has only seldom been analyzed in this species and its meaning is still not fully understood. It has been suggested that a
high FES may reflect a strongly piscivorous behaviour while low FES could reflect a higher utilization of invertebrates. We
compared the stomach contents and the trophic position of northern pike in 16 populations of individuals feeding mainly on
fish or benthic invertebrates. We tested the hypothesis that northern pike with empty stomachs or with fish in their stomachs
have a higher trophic position than individuals feeding on invertebrates. Carbon (δ13C) and nitrogen (δ15N) stable isotope signatures were used to estimate the trophic position of individuals. We found no significant difference
in the trophic position among piscivores, invertebrate feeders, and northern pike with empty stomachs. The average trophic
position of northern pike was high (mean ± SD = 4.3 ± 0.4, n = 66) and was correlated with total length. These results indicate that, although invertebrates could be an important part
of the diet of northern pike in Canadian Shield lakes, fish are still the dominant prey. Hence, feeding on invertebrates in
our study lakes would reflect an opportunistic rather than a specialized feeding strategy. 相似文献
Prostaglandins (PGs) are lipid mediators that may play important roles in cancer, immunomodulation, and neurodegeneration. So, the quantitative analysis of PGs will therefore be important in order to understand the natural history of a range of diseases and may be used as a tool in the development of new biotherapeutics. However, such an analysis is problematic because of the small quantities of PGs present in the body. Here, we developed a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-MS)-based analytical method for rapid and easy quantification of the ketone-containing PGs (15d-PGJ2etc.) as a targeted metabolomics platform. The chemical derivatization with Girard’s reagent P (GP) provided a good linearity (R2 > 0.99) between peak area and quantity of 15d-PGJ2 and highly improved sensitivity (limit of quantitation, LOQ: 1.25 pmol on a spot in MALDI plate). Finally, we utilized this method to directly characterize the interaction between peroxisome proliferatoractivated receptor gamma-ligand binding domain (PPARγ-LBD) and 15d-PGJ2. The 15d-PGJ2 was enriched by PPARγ-LBD and also the binding level of the ligand was dropped considerably by the treatment of PPARγ agonist such as rosiglitazone (about 0.61-fold reduction). Taken together, our MALDI-MS-based targeted metabolomics method for ketone-containing PGs may be applicable to elucidate the protein-metabolite interactions and to identify natural ligands for drug candidates. 相似文献
Paramagnetic relaxation enhancements (PRE) present a powerful source of structural information in nuclear magnetic resonance (NMR) studies of proteins and protein–ligand complexes. In contrast to conventional PRE reagents that are covalently attached to the protein, the complex between gadolinium and three dipicolinic acid (DPA) molecules, [Gd(DPA)3]3?, can bind to proteins in a non-covalent yet site-specific manner. This offers straightforward access to PREs that can be scaled by using different ratios of [Gd(DPA)3]3? to protein, allowing quantitative distance measurements for nuclear spins within about 15 Å of the Gd3+ ion. Such data accurately define the metal position relative to the protein, greatly enhancing the interpretation of pseudocontact shifts induced by [Ln(DPA)3]3? complexes of paramagnetic lanthanide (Ln3+) ions other than gadolinium. As an example we studied the quaternary structure of the homodimeric GCN4 leucine zipper. 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.
The cystine/glutamate exchanger (antiporter xc−) is a membrane transporter involved in the uptake of cystine, the rate-limiting amino acid in the synthesis of glutathione.
Recent studies suggest that the antiporter plays a role in the slow oxidative excitotoxity and in the pathological effects
of β-N-oxalylamino-l-alanine, the molecule responsible for neurolathyrism, a neurotoxic upper motor neuron disease. The mouse cystine/glutamate
exchanger has been cloned and showed to be composed of two distinct proteins, one of which being a novel protein, named xCT,
of 502 amino acids and 12 putative trans-membrane domains. We have generated and purified a polyclonal antibody to mouse xCT
and studied its expression in rat brain and in different cultured cells (astrocytes, fibroblasts and neurons) using Western
blot and immunocytochemical techniques. Expression of xCT was also studied in rat brain and muscle at different developmental
stages. Parallel experiments were carried out with antibodies to the heavy chain of 4F2 surface antigen, the non-specific
subunit of the antiporter xc−. xCT antibody detected in all cell and tissue extracts a specific band of about 40 kDa. Subcellular fractionation demonstrated
that xCT is concentrated mainly in the microsomal-mitochondrial fraction, in accord with its structure as transmembrane protein.
Immunocytochemical analysis showed a strong staining in all cells examined, included neurons. Furthermore, both xCT and the
heavy chain of 4F2 surface antigen increased in the brain during development, reaching the highest expression in adulthood.
The study of the expression and developmental profile of xCT represents a first step towards a better characterization of
its biochemical properties and function, which in turn may help to understand the relative contribution of the xc− antiporter in the pathogenesis of certain neurodegenerative diseases. 相似文献
Тhe DNA testing of autosomal recessive deafness type 1A (DFNB1A, MIM 220290) is complicated when deaf patients have only monoallelic (heterozygous) recessive mutations in the GJB2 (Сх26) gene that is uninformative for establishment of diagnosis. Such patients may be “random” heterozygous carriers of GJB2 mutations as well as have the mutant allele in a cis-regulatory region of GJB2 gene, in element genes encoding other connexins: GJB6 (Сх30) or GJB3 (Сх31). Previous studies of genetic causes of hearing loss in patients from Yakutia were directed to search for only mutations in the GJB2 gene, and the DNA diagnostics was uninformative for 9.7% (38/393) of the patients with monoallelic GJB2 mutations. In this work the search for mutations in genes GJB3 and GJB6 and two deletions с.del(GJB6-D13S1830) and с.del(GJB6-D13S1854) to the cis-regulatory region of GJB2 gene was conducted in 35 patients with GJB2 monoallelic mutations and in 104 normal hearing individuals. The genes studied are two synonymous substitution c.489G>A (р.Leu163Leu) (GJB6) and c.357C>T (р.Asn119Asn) (GJB3) have been found, probably do not have clinical significance, and two nonsynonymous substitution c.301G>A (p.Glu101Lys) (GJB6) and с.580G>A (p.Ala194Thr) (GJB3). Additional experimental evidences are needed for confirmation of pathogenic significance of detected nonsynonymous substitutions in development of hearing loss in studied patients. Diagnosis of the DFNB1A was confirmed in only one patient, who was discovered by the deletion с.del(GJB6-D13S1830) (GJB2) in combination with a recessive mutation с.35delG (GJB2). In general, our results indicate low contribution of mutations in genes GJB6 and GJB3 in hearing loss etiology in Yakutia. 相似文献
Among various types of ionizing radiation, the beta emitter radionuclides are involved in many sectors of human activity, such as nuclear medicine, nuclear industries and biomedicine, with a consequently increased risk of accidental, occupational or therapeutic exposure. Despite their recognized importance, there is little information about the effect of beta particles at the cellular level when compared to other types of ionizing radiation. Thus, the objective of the present study was to evaluate the genotoxic and cytotoxic effects of 90Sr/90Y—a pure, highly energetic beta source—on Chinese hamster ovary (CHO) cells and to compare them with data obtained with 60Co. CHO cells irradiated with different doses of 60Co (0.34 Gy min–1) and 90Sr/90Y (0.23 Gy min–1) were processed for analysis of clonogenic death, induction of micronuclei (MN) and interphase death. The survival curves obtained for both types of radiation were fitted by the exponential quadratic model and were found to be similar. Also, the cytogenetic results showed similar frequencies of radio-induced MN between gamma and beta radiations and the MN distribution pattern among cells did not follow the expected Poisson probability pattern. The relative variance values were significantly higher in cells irradiated with 90Sr/90Y than with 60Co in all exposure doses. The irradiated cells showed more necrotic cells 72 h and 96 h after exposure to beta than to gamma radiation. In general, the 90Sr/90Y -radiation was more damaging than 60Co -rays. The data obtained also demonstrated the need to use several parameters for a better estimate of cellular sensitivity to the action of genotoxic agents, which would be important in terms of radiobiology, oncology and therapeutics. 相似文献
The nonenzymatic isoprostane pathway of lipid peroxidation of polyunsaturated fatty acids results in formation of products, termed isoprostanes, which have very large positional and stereo isomerism, possess various biological activities, produce adducts with proteins, and thus contribute to pathogeneses of the agedependent diseases. However, it was unclear what mechanism drives this type of lipid autoxidation, and why the products have very large isomerism. We propose a mechanism when perhydroxyl radicals (HO2?) react with polyunsaturated fatty acids in the hydrophobic milieu of membranes. In the membrane HO2? initiates a chain of reactions with formation first H2O2, which undergoes homolytic fission producing two ?OH radicals, thus very rapidly abstracting three H atoms from a polyunsaturated fatty acid. As a result, the HO2? molecule is converted to two molecules of water, and the molecule of a polyunsaturated fatty acid loses two double bonds, becomes highly unstable and undergoes peroxidation and random intramolecular re-arrangements causing a very large isomerism of the final products. The extremely high reactivity of ?HО2 with polyunsaturated fatty acids is the cause of very subtle and slow accumulation of damages in the membrane and membrane associated proteins, even though the concentration of ?HО2 relative to superoxide radical may be very low. 相似文献
Alkaline-earth metal complexes of the monoanionic form of the polyether ionophore monensin A were isolated for the first time
in solid state and were structurally characterized using various spectroscopic methods (IR, NMR, FAB-MS). The stoichiometric
reaction of monensic acid (MonH) with M2+ (M = Mg, Ca) in the presence of an organic base leads to the formation of mononuclear complexes of composition [M(Mon)2(H2O)2]. The structures of magnesium (1) and calcium (2) monensin complexes in the solid state were established by single crystal X-ray crystallography. The complexes crystallize
as [Mg(Mon)2(H2O)2]·5MeCN (1) and [Ca(Mon)2(H2O)2]·H2O·5MeCN (2) in the monoclinic P21 space group. The alkaline-earth metal ion is placed in a distorted octahedral environment, defined by two monensin anions
acting as bidentate ligands in the equatorial plane of the complex as well as by two water molecules occupying the axial positions
of the inner coordination sphere. The bactericidal activity of 1 and 2 was evaluated against aerobic Gram-positive microorganisms applying the double layer agar hole diffusion method. 相似文献
An extension to HN(CO-α/β-N,Cα-J)-TROSY (Permi and Annila in J Biomol NMR 16:221–227, 2000) is proposed that permits the simultaneous determination of the four coupling constants 1JN′(i)Cα(i), 2JHN(i)Cα(i), 2JCα(i−1)N′(i), and 3JCα(i−1)HN(i) in 15N,13C-labeled proteins. Contrasting the original scheme, in which two separate subspectra exhibit the 2JCαN′ coupling as inphase and antiphase splitting (IPAP), we here record four subspectra that exhibit all combinations of inphase
and antiphase splittings possible with respect to both 2JCαN′ and 1JN′Cα (DIPAP). Complementary sign patterns in the different spectrum constituents overdetermine the coupling constants which can
thus be extracted at higher accuracy than is possible with the original experiment. Fully exploiting data redundance, simultaneous
2D lineshape fitting of the E.COSY multiplet tilts in all four subspectra provides all coupling constants at ultimate precision.
Cross-correlation and differential-relaxation effects were taken into account in the evaluation procedure. By applying a four-point
Fourier transform, the set of spectra is reversibly interconverted between DIPAP and spin-state representations. Methods are
exemplified using proteins of various size. 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
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
A confined aquifer in the Malm Karst of the Franconian Alb, South Germany was investigated in order to understand the role of the vadose zone in denitrifiaction processes. The concentrations of chemical tracers Sr2+ and Cl– and concentrations of stable isotope 18O were measured in spring water and precipitation during storm events. Based on these measurements a conceptual model for runoff was constructed. The results indicate that pre-event water, already stored in the system at the beginning of the event, flows downslope on vertical and lateral preferential flow paths. Chemical tracers used in a mixing model for hydrograph separation have shown that the pre-event water contribution is up to 30%. Applying this information to a conceptual runoff generation model, the values of 15N and 18O in nitrate could be calculated. Field observations showed the occurence of significant microbial denitrification processes above the soil/bedrock interface before nitrate percolates through to the deeper horizon of the vadose zone. The source of nitrate could be determined and denitrification processes were calculated. Assuming that the nitrate reduction follows a Rayleigh process one could approximate a nitrate input concentration of about 170 mg/l and a residual nitrate concentration of only about 15%. The results of the chemical and isotopic tracers postulate fertilizers as nitrate source with some influence of atmospheric nitrate. The combined application of hydrograph separation and determination of isotope values in 15N and 18O of nitrate lead to an improved understanding of microbial processes (nitrification, denitrification) in dynamic systems. 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
Oxidative stimulation induced by hydrogen peroxide (H2O2) on human lens epithelial cells (HLECs) was performed to observe the effects on cell viability and caveolin expression, and
cholesterol depletion in HLECs caused by methyl-β-cyclodextrin (MβCD) was also studied. SRA01/04 HLECs were exposed to H2O2 or MβDC of various concentrations and durations. We used a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)
assay to measure the effect of H2O2 on the viability of SRA01/04 HLECs. The distributions of caveolins after oxidative stimulation were probed by fluorescence
microscopy and laser scanning confocal microscopy. Immunoblotting was performed to analyze alterations of caveolins expression.
We observed that the viability of SRA01/04 HLECs under 0.1 mM H2O2 for 10 min or longer was significantly reduced (*p < 0.05, F = 11.63). Laser scanning microscopy showed immunofluorescent caveolins in SRA01/04 HLECs under 0.1 mM H2O2 for 10 min or longer; caveolins were largely confined to intracellular domains. Western blots showing both membrane and total
caveolin protein (22 kDa) levels in SRA01/04 HLECs treated with 0.1, 0.2, 0.5, or 1.0 mM H2O2 for 30 min were significantly reduced, compared with the untreated (*p < 0.05, F = 6.149, or *p < 0.05, F = 14.489, respectively). In addition, the membrane and total caveolin protein level after being treated with 0.1 mM H2O2 (*p < 0.05, F = 6.843, or *p < 0.05, F = 7.944, respectively) for different durations also down regulated. Fluorescence microscopy also showed that phosphorylated
caveolin-1 was distributed near the focal adhesions of the cells. This study concludes that the responses of HLECs to oxidative
stress may include down regulation of caveolin and phosphorylation of caveolin-1 on tyrosine 14 and that MβCD also down regulates
caveolin while depleting cholesterol in HLECs.
Published in Russian in Molekulyarnaya Biologiya, 2007, Vol. 41, No. 6, pp. 994–1001.
The article was submitted by the authors in English. 相似文献
While tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is a promising new agent for the treatment of
cancer, resistance to TRAIL remains a therapeutic challenge. Identifying agents to use in combination with TRAIL to enhance
apoptosis in leukemia cells would increase the potential utility of this agent as a therapy for leukemia. Here, we show that
15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2), a natural ligand for peroxisome proliferator-activated receptor γ (PPARγ), can sensitize TRAIL-resistant leukemic HL-60
cells to TRAIL-induced apoptosis. The sensitization to TRAIL-induced apoptosis by 15d-PGJ2 was not blocked by a PPARγ inhibitor (GW9662), suggesting a PPARγ-independent mechanism. This process was accompanied by
activation of caspase-8, caspase-9, and caspase-3 and was concomitant with Bid and PARP cleavage. We observed significant
decreases in XIAP, Bcl-2, and c-FLIP after cotreatment with 15d-PGJ2 and TRAIL. We also observed the inhibition of Akt expression and phosphorylation by cotreatment with 15d-PGJ2 and TRAIL. Furthermore, inactivation of Akt by Akt inhibitor IV sensitized human leukemic HL-60 cells to TRAIL, indicating
a key role for Akt inhibition in these events. Taken together, these findings indicate that 15d-PGJ2 may augment TRAIL-induced apoptosis in human leukemia cells by down-regulating the expression and phosphorylation of Akt. 相似文献
