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1.
Madoka Takahashi Wakako Handa Hiromi Umeta Saki Ishikawa Kouwa Yamashita Mitsuteru Numazawa 《The Journal of steroid biochemistry and molecular biology》2009,116(3-5):191-199
To gain the structure–activity relationship of Δ1-androstenediones (Δ1-ADs) as mechanism-based inactivator of aromatase, series of 2-alkyl- and 2-alkoxy-substitiuted Δ1-ADs (6 and 9) as well as 2-bromo-Δ1-AD (14) were synthesized and tested. All of the inhibitors examined blocked aromatase in human placental microsomes in a competitive manner. In a series of 2-alkyl-Δ1-ADs (6), n-hexyl compound 6f was the most powerful inhibitor with an apparent Ki value of 31 nM. The inhibitory activities of 2-alkoxy steroids 9 decreased in relation to length of the alkyl chain up to n-hexyloxy group (Ki: 95 nM for methoxy 9a). All of the alkyl steroids 6 along with the alkoxy steroid 9, except for the ethyl and n-propyl compounds 6b and 6c, caused a time-dependent inactivation of aromatase. The inactivation rates (kinact: 0.020–0.084 min−1) were comparable to that of the parent compound Δ1-AD. The inactivation was prevented by the substrate AD, and no significant effect of l-cysteine on the inactivation was observed in each case. The results indicate that the 2-hexyl compound 6f act as the most powerful mechanism-based inactivator of aromatase among Δ1-AD analogs and may be submitted to the preclinical study in estrogen-dependent breast cancer. 相似文献
2.
The effects of E2 levuglandins on the contractile activity of rat uterine horns were studied. LGE2, AnLGE2, delta 9-LGE2 and the synthetic epimer, 8-epi-delta 9-LGE2 all induced contractions in a dose-response fashion. AnLGE2 gave decreased responses with increased bath concentrations. Paired comparisons showed potent and selective inhibitory effects of AnLGE2 on the uterotonic activity of prostaglandins. AnLGE2 inhibited the uterotonic activity of PGE2 at a 0.1:1 ratio, of PGD2 at a 1:1 ratio, but did not inhibit the activity of PGF2 alpha. Exposure of spontaneously contracting uteri to high concentrations of AnLGE2, or prolonged exposure to lower concentrations, suppressed contractions. 相似文献
3.
P. Sirois S. Roy G. Beraldin P. Valler P. Borgeat 《Prostaglandins & other lipid mediators》1982,24(3)
The activity of natural 5,6-Dihydroxy-eicosatetraenoic acid (5,6-DiHETE; 2 isomers), 5S,15S-DiHETE, 8S,15S-DiHETE, 5S,12S-DiHETE, Δ6-trans-leukotriene B4, 12-epi-Δ6-leukotriene B4, ω-hydroxy-leukotriene B4, ω-carboxy-leukotriene B4, 15S-hydroxy-eicosatetraenoic acid (15S-HETE), 12S-HETE, 5S-HETE and 12S-hydroxy-heptadecatrienoic acid was compared to TLB4 on the guinea-pig lung parenchymal strip and on the release of prostaglandins and thromboxanes by the perfused guinea-pig lungs. The ω-hydroxy-LTB4 appeared more potent than LTB4 both for inducing a contraction and for releasing prostanoids whereas the ω-carboxy-LTB4 was much less active on the parenchyma and did not release prostanoids at the dose used. All other hydroxy acids tested were either very weakly active or inactive in the two systems used with the exception of the 5,6-DiHETEs which showed significant activity. These di-hydroxy acids induced contractions of the lung parenchymal strip which could be blocked by PFL-55712 but were inactive on the guinea-pig ileum. The 5S-HETE, 12S-HETE and 15S-HETE were also tested for possible myotropic activity on selected smooth muscle preparations. Our results provide further informations on the structural requirements for LTB4 (and other hydroxy acids) actions on the guinea-pig lungs. 相似文献
4.
We investigate the effect of the prostaglandin D2 metabolite Δ12−PGD2 (9−Deoxy−Δ9, Δ12−13,14-dihydroprostaglandin D2) on collagen synthesis in human osteoblast. Δ12-PGJ2 at 10−5M enhanced collagen synthesis in the presence of 2 mM α-glycerophosphate-2Na. The stimulative effect appeared as early as 3 days after addition and continued until 22 days. The enhancement of type I collagen synthesis was confirmed by polyacrylamide gel electrophoresis. The potency was the same as 101t-8M 1 α, 25 dihydroxy vitamine D3 (1,25(OH)2D3). Northern blot analysis showed that 10−5M Δ 12-PGD2 and 10−8M 1,25(OH)2D3 enhanced the transcribtion of type 1 procollagen (α1) mRNA levels in osteoblasts. 相似文献
5.
Levuglandin E2 (LGE2), a γ-ketoaldehyde produced by rearrangement of the prostaglandin endoperoxide PGH2 under the aqueous conditions of its biosynthesis, causes extensive intermolecular crosslinking of ovalbumin at pH 6 or pH 7 and 37°C. The time dependence of protein oligomerization is monitored by SDS-PAGE. Effects of pH and concentration on the extent of LGE2-induced crosslinking are examined. The efficacy of LGE2 for inducing crosslinking is compared with other oxidative metabolites of arachidonic acid (AA), including the prostaglandins PGE2, PGD2, PGA2, PGB2, and PGF2α, as well as malondialdehyde and E-4-hydroxy-non-2-enal. LGE2 is orders of magnitude more effective in crosslinking protein than any other cyclooxygenase or lipoxygenase metabolite of AA tested. 相似文献
6.
Yeast cytochrome c peroxidase (CCP) efficiently catalyzes the reduction of H2O2 to H2O by ferrocytochrome c in vitro. The physiological function of CCP, a heme peroxidase that is targeted to the mitochondrial intermembrane space of Saccharomyces cerevisiae, is not known. CCP1-null-mutant cells in the W303-1B genetic background (ccp1Δ) grew as well as wild-type cells with glucose, ethanol, glycerol or lactate as carbon sources but with a shorter initial doubling time. Monitoring growth over 10 days demonstrated that CCP1 does not enhance mitochondrial function in unstressed cells. No role for CCP1 was apparent in cells exposed to heat stress under aerobic or anaerobic conditions. However, the detoxification function of CCP protected respiring mitochondria when cells were challenged with H2O2. Transformation of ccp1Δ with ccp1W191F, which encodes the CCPW191F mutant enzyme lacking CCP activity, significantly increased the sensitivity to H2O2 of exponential-phase fermenting cells. In contrast, stationary-phase (7-day) ccp1Δ-ccp1W191F exhibited wild-type tolerance to H2O2, which exceeded that of ccp1Δ. Challenge with H2O2 caused increased CCP, superoxide dismutase and catalase antioxidant enzyme activities (but not glutathione reductase activity) in exponentially growing cells and decreased antioxidant activities in stationary-phase cells. Although unstressed stationary-phase ccp1Δ exhibited the highest catalase and glutathione reductase activities, a greater loss of these antioxidant activities was observed on H2O2 exposure in ccp1Δ than in ccp1Δ-ccp1W191F and wild-type cells. The phenotypic differences reported here between the ccp1Δ and ccp1Δ-ccp1W191F strains lacking CCP activity provide strong evidence that CCP has separate antioxidant and signaling functions in yeast. 相似文献
7.
The kinetics of substitution reactions of [η-CpFe(CO)3]PF6 with PPh3 in the presence of R-PyOs have been studied. For all the R-PyOs (R = 4-OMe, 4-Me, 3,4-(CH)4, 4-Ph, 3-Me, 2,3-(CH)4, 2,6-Me2, 2-Me), the reactions yeild the same product [η5-CpFe(CO)2PPh3]PF6, according to a second-order rate law that is first order in concentrations of [η5-CpFe(CO)3]PF6 and of R-PyO but zero order in PPh3 concentration. These results, along with the dependence of the reaction rate on the nature of R-PyO, are consistent with an associative mechanism. Activation parameters further support the bimmolecular nature of the reactions: ΔH≠ = 13.4 ± 0.4 kcal mol−1, ΔS≠ = −19.1 ± 1.3 cal k−1 mol−1 for 4-PhPyO; ΔH≠ = 12.3 ± 0.3 kcal mol−1, ΔS≠ = 24.7 ±1.0 cal K−1 mol−1 for 2-MePyO. For the various substituted pyridine N-oxides studied in this paper, the rates of reaction increase with the increasing electron-donating abilities of the substituents on the pyridine ring or N-oxide basicities, but decrease with increasing 17O chemical shifts of the N-oxides. Electronic and steric factors contributing to the reactivity of pyridine N-oxides have been quantitatively assessed. 相似文献
8.
The pitching-down flapping is a new type of bionic flapping,which was invented by the author based on previous studieson the aerodynamic mechanisms of fruit fly(pitching-up)flapping.The motivation of this invention is to improve the aerodynamiccharacteristics of flapping Micro Air Vehicles(MAVs).In this paper the pitching-down flapping is briefly introduced.Themajor works include:(1)Computing the power requirements of pitching-down flapping in three modes(advanced,symmetrical,delayed),which were compared with those of pitching-up flapping;(2)Investigating the effects of translational accelerationtime,Δτ_t,and rotational time,Δτ_r,at the end of a stroke,and the angle of attack,α,in the middle of a stroke on the aerodynamiccharacteristics in symmetrical mode;(3)Investigating the effect of camber on pitching-down flapping.From the above works,conclusions can be drawn that:(1)Compared with the pitching-up flapping,the pitching-down flapping can greatly reduce thetime-averaged power requirements;(2)The increase in Δτt and the decrease in Δτ_r can increase both the lift and drag coefficients,but the time-averaged ratio of lift to drag changes a little.And α has significant effect on the aerodynamic characteristicsof the pitching-down flapping;(3)The positive camber can effectively increase the lift coefficient and the ratio of lift to drag. 相似文献
9.
John W. Huffman John Liddle Shu Yu Mie Mie Aung Mary E. Abood Jenny L. Wiley Billy R. Martin 《Bioorganic & medicinal chemistry》1999,7(12):964
The synthesis and pharmacology of 15 1-deoxy-Δ8-THC analogues, several of which have high affinity for the CB2 receptor, are described. The deoxy cannabinoids include 1-deoxy-11-hydroxy-Δ8-THC (5), 1-deoxy-Δ8-THC (6), 1-deoxy-3-butyl-Δ8-THC (7), 1-deoxy-3-hexyl-Δ8-THC (8) and a series of 3-(1′,1′-dimethylalkyl)-1-deoxy-Δ8-THC analogues (2, n=0–4, 6, 7, where n=the number of carbon atoms in the side chain−2). Three derivatives (17–19) of deoxynabilone (16) were also prepared. The affinities of each compound for the CB1 and CB2 receptors were determined employing previously described procedures. Five of the 3-(1′,1′-dimethylalkyl)-1-deoxy-Δ8-THC analogues (2, n=1–5) have high affinity (Ki=<20 nM) for the CB2 receptor. Four of them (2, n=1–4) also have little affinity for the CB1 receptor (Ki=>295 nM). 3-(1′,1′-Dimethylbutyl)-1-deoxy-Δ8-THC (2, n=2) has very high affinity for the CB2 receptor (Ki=3.4±1.0 nM) and little affinity for the CB1 receptor (Ki=677±132 nM).
Scheme 3. (a) (C6H5)3PCH3+ Br−, n-BuLi/THF, 65°C; (b) LiAlH4/THF, 25°C; (c) KBH(sec-Bu)3/THF, −78 to 25°C then H2O2/NaOH. 相似文献
10.
Statistical analysis reveals that the set of differences between the secondary shifts of the α- and β-carbons for residues i of a protein (Δδ13Cαi- Δδ13Cβi) provides the means to detect and correct referencing errors for 1H and 13C nuclei within a given dataset. In a correctly referenced protein dataset, linear regression plots of Δδ13Cαi,Δδ13Cβi, or Δδ1Hαi vs. (Δδ13Cαi- Δδ13Cβi) pass through the origin from two directions, the helix-to-coil and strand-to-coil directions. Thus, linear analysis of chemical shifts (LACS) can be used to detect referencing errors and to recalibrate the 1H and 13C chemical shift scales if needed. The analysis requires only that the signals be identified with distinct residue types (intra-residue spin systems). LACS allows errors in calibration to be detected and corrected in advance of sequence-specific assignments and secondary structure determinations. Signals that do not fit the linear model (outliers) deserve scrutiny since they could represent errors in identifying signals with a particular residue, or interesting features such as a cis-peptide bond. LACS provides the basis for the automated detection of such features and for testing reassignment hypotheses. Early detection and correction of errors in referencing and spin system identifications can improve the speed and accuracy of chemical shift assignments and secondary structure determinations. We have used LACS to create a database of offset-corrected chemical shifts corresponding to nearly 1800 BMRB entries: 300 with and
1500 without corresponding three-dimensional (3D) structures. This database can serve as a resource for future analysis of the effects of amino acid sequence and protein secondary and tertiary structure on NMR chemical shifts.Supplementary material to this paper is available in electronic form at
http://dx.doi.org/10.1007/s10858-005-1717-0 相似文献
11.
We studied the effect of intracellular glutathione (GSH), which was known to conjugate readily with an α, β-unsaturated carbonyl of 9-deoxy-Δ9,12-13,14-dihydro PGD2 (Δ12-PGJ2), on the cytotoxicity of Δ12-PGJ2. Δ12-PGJ2 caused DNA fragmentation in human hepatocellular carcinoma Hep 3B cells, which was blocked by cycloheximide (CHX). The Δ12-PGJ2-induced apoptosis was augmented by GSH depletion resulted from pretreatment with buthioninine sulfoximine (BSO), an inhibitor of γ-glutamylcysteine synthetase. On the contrary, N-acetyl-cysteine (NAC), a precursor of cysteine, elevated the GSH level and protected cells from initiating apoptosis by Δ12-PGJ2. Sodium arsenite, a thiol-reactive agent, also induced apoptosis, which was potentiated or attenuated by BSO or NAC treatment respectively. These results suggest that the apoptosis-inducing activity of Δ12-PGJ2 is due to thiol-reactivity and intracellular GSH modulates the Δ12-PGJ2-induced apoptosis by regulating the accessibility of Δ12-PGJ2 to target proteins containing thiol groups. 相似文献
12.
Hiroya Tanaka Takao Yamamuro Manabu Matsumoto Yoshihiko Kotoura Chiaki Tanaka 《Prostaglandins & other lipid mediators》1985,30(1)
Effects of antineoplastic prostaglandins (PG), PGD2 and 9-deoxy-Δ9-PGD2, on colony formation of cloned Dunn osteosarcoma (TA 102), normal Swiss 3T3 and V-79 cell lines were evaluated. PGD2 significantly inhibited the colony formation of TA 102 cells in a dose-dependent manner at concentrations between 0.5 and 5 ug/ml. The IC50 value was calculated to be 0.72 ug/ml. A dose-dependent inhibition of TA 102 colony formation was also observed with 9-deoxy-Δ9-PGD2 between 0.01 to 1 ug/ml, the IC50 value being 0.22 ug/ml. These prostaglandins did not exert cytocidal effects in vitro on Swiss 3T3 cells at concentrations between 0.01 to 1 ug/ml. The two agents had no significant cytocidal effects on V-79 cells except for 9-deoxy-Δ9-PGD2 at a concentration of 5 ug/ml. These results suggest that PGD2 and 9-deoxy-Δ9-PGD2 are considered to have cytocidal activity on Dunn osteosarcoma cells in dosages which do not affect non-malignant cells. 相似文献
13.
OCP1 and OCP2, the most abundant proteins in the cochlea, are evidently subunits of an SCF E3 ubiquitin ligase. Although transcribed from a distinct gene, OCP2 is identical to Skp1. OCP1 is equivalent to the F-box protein known as Fbs1, Fbx2, or NFB42 — previously shown to bind N-glycosylated proteins and believed to function in the retrieval and recycling of misfolded proteins. The high concentrations of OCP1 and OCP2 in the cochlea suggest that the OCP1–OCP2 heterodimer may serve an additional function independent of its role in a canonical SCF complex. At 25 °C, urea-induced denaturation of OCP1 is slow, but reversible. The data suggest that the protein possesses one or more disordered regions, a conclusion supported by analysis of the far-UV circular dichroism spectrum and the appearance of the 1H, 15N-HSQC spectrum. Thermal denaturation of OCP1 is irreversible, evidently due to formation of high molecular weight aggregates. Analysis with a kinetic model yields an estimate for the activation energy for unfolding of 49 kcal/mol. Urea denaturation data for OCP2 returns ΔGo and m values of 6.2 kcal/mol and 1.5 kcal mol− 1 M− 1, respectively. In contrast to OCP1, thermal denaturation of OCP2 is reversible. In phosphate-buffered saline, at pH 7.40, the protein exhibits a ΔHvH/ΔHcal ratio of 1.69, suggesting that denaturation proceeds largely from the native dimer directly to the unfolded state. OCP1 and OCP2 associate tightly at room temperature. However, DSC data for the complex suggest that they denature independently, consistent with the highly exothermic enthalpy of complex formation reported previously. 相似文献
14.
John W. Huffman Simon M. Bushell John R. A. Miller Jenny L. Wiley Billy R. Martin 《Bioorganic & medicinal chemistry》2002,10(12):7809
Three series of new cannabinoids were prepared and their affinities for the CB1 and CB2 cannabinoid recptors were determined. These are the 1-methoxy-3-(1′,1′-dimethylalkyl)-, 1-deoxy-11-hydroxy-3-(1′,1′-dimethylalkyl)- and 11-hydroxy-1-methoxy-3-(1′,1′-dimethylalkyl)-Δ8-tetrahydrocannabinols, which contain alkyl chains from dimethylethyl to dimethylheptyl appended to C-3 of the cannabinoid. All of these compounds have greater affinity for the CB2 receptor than for the CB1 receptor, however only 1-methoxy-3-(1′,1′-dimethylhexyl)-Δ8-THC (JWH-229, 6e) has effectively no affinity for the CB1 receptor (Ki=3134±110 nM) and high affinity for CB2 (Ki=18±2 nM). 相似文献
15.
Di-nor-benzofuran neolignan aldehydes, Δ7-3,4-methylenedioxy-3′-methoxy-8′,9′-dinor-4′,7-epoxy-8,3′-neolignan-7′-aldehyde (ocophyllal A) 1, Δ7-3,4,5,3′-tetramethoxy-8′,9′-dinor-4′,7-epoxy-8,3′-neolignan-7′-aldehyde (ocophyllal B) 2, and macrophyllin-type bicyclo[3.2.1]octanoid neolignans (7R, 8R, 3′S, 4′S, 5′R)-Δ8′-4′-hydroxy-5′-methoxy-3,4-methylenedioxy-2′,3′,4′,5′-tetrahydro-2′-oxo-7.3′,8.5′-neolignan (ocophyllol A) 3, (7R, 8R, 3′S, 4′S, 5′R)-Δ8′-4′-hydroxy-3,4,5′-trimethoxy-2′,3′,4′,5′-tetrahydro-2′-oxo-7.3′,8.5′-neolignan (ocophyllol B) 4, (7R, 8R, 3′S, 4′S, 5′R)-Δ8′-4′-hydroxy-3,4,5,5′-tetramethoxy-2′,3′,4′,5′-tetrahydro-2′-oxo-7.3′,8.5′-neolignan (ocophyllol C) 5, as well as 2′-epi-guianin 6 and (+)-licarin B 7, were isolated and characterized from leaves of Ocotea macrophylla (Lauraceae). The structures and configuration of these compounds were determined by extensive spectroscopic analyses. Inhibition of platelet activating factor (PAF)-induced aggregation of rabbit platelets were tested with neolignans 1–7. Although compound 6 was the most potent PAF-antagonist, compounds 3–5 showed some activity. 相似文献
16.
Elizabeth M.K. Leovey Niels H. Andersen Pamela Bissonette 《Prostaglandins & other lipid mediators》1979,17(1)
, originally introduced as an inadvertent contaminant in solutions used for evaluating the stability of prostaglandins, proved to lead to the rapid disappearance of the cyclopentenone unit of PGA2 (as monitored by circular dichroic spectroscopy). The cyclopentenone unit is converted, in various metabolites, to a 9-keto, 9α or 9β-hydroxy group lacking the ring unsaturation. The major EtoAc-soluble 9-hydroxy metabolite (Compound-I) was shown to be 9α, 15α-dihydroxy-2,3,4,5-tetranor-13-
-prostenoic acid. Similar tetranor 9-hydroxy metabolites with one additional degree of unsaturation, and with a 9β-hydroxy group, also occur but these have not been fully characterized. Only two of the wide range of 9-keto metabolites are fully characterized by mass spectral (MS) data: 9,15-oxo-2,3,4,5-tetranorprostanoic acid and 9,15-oxo-2,3,4,5-tetranor-13-
-prostenoic acid. The water soluble metabolites have not been characterized further.The fully characterized metabolites together with MS data from mixtures of minor metabolites indicate that
can perform the following transformations: β-oxidation, dehydrogenation at C-15, reduction of the enone carbon-carbon double bonds (both Δ10,11 and Δ13,14), reduction of the 9-ketone, and possibly migration of the cyclopentyl double bond (Δ10,11 → Δ11,12).
metabolizes 15-epimeric PGA2 equally readily with the production of similar products. PGA1 affords less 9-keto metabolites with compound I constituting 33% of the product by HPLC analysis.
displays some enantioselectivity, PGA2 and 15-epi-PGA2 are each metabolized more rapidly than their enantiomers. Other prostaglandins appear to be less readily metabolized. 相似文献
17.
Yoshitaka Shimizu Shinjiro Todo Shinsaku Imashuku 《Prostaglandins & other lipid mediators》1986,32(4)
To study the precise mechanism of cytotoxic activity of PGD2 or Δ12-PGJ2 (a biological active metabolite of PGD2), we examined the effect of various compounds on PGD2 or Δ12-PGJ2 cytottoxic, using a human neuroblastoma cell line (NCG). Cycloheximide (CHM) specifically protected PGD2 cytotoxicity on NCG cells. When Δ12-PGJ2 was tested, CHM exhibited a similar rescue effect. Puromycin, mitomycin C, and α-amanitin did not affect PGD2 or Δ12-PGJ2 cytotoxicity. Emetine showed a variable and no consistent rescue effect CHM may have been active at the primary site where PGD2 or Δ12-PGJ2 exerts its cytotoxicity. This is the first report indicating that CHM reduces the cytotoxicity induced by PGD2 or Δ12-PGJ2. 相似文献
18.
Jeak Ling Ding Yong Zhu Bow Ho 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2001,759(2):237-246
Synthetic peptide S3Δ has high affinity for bacterial endotoxin or lipopolysaccharide (LPS). Under tested conditions of pH 5–9 and 0–0.4 M NaCl, the affinity constant, KD ranged from 2·10−6 to 2·10−9 M−1. A novel affinity matrix based on peptide S3Δ was developed for removal of LPS from solutions such as: water; buffers with a wide range of ionic strength and pH; medium for cell culture; and protein solutions under optimized conditions. At a starting LPS of ≈100 EU/ml, a post-purification level below 0.005 EU/ml was achieved. 相似文献
19.
PGJ2 and Δ12PGJ2 (1 μM to 30 μm) inhibited the growth of human astrocytoma cells (1321N1) in a time-dependent manner within 48 hrs, determined by [3H]thymidine incorporation into acid-insoluble fraction or amounts of protein. The EC50 values for PGJ2 and Δ12PGJ2 were approximately 8 μM and 6 μM, respectively. [3H]Thymidine incorporation to acid insoluble fraction was inhibited by these PGs within 1 hr, indicating that these PGs rapidly affect cell functions. Although it has been reported that an increase in cyclic AMP inhibits cell growth, PGJ2 and Δ12PGJ2, but not PGE1, reduced isoproterenol (10 μM)-induced accumulation of cyclic AMP, suggesting that PGJ2 and Δ12PGJ2 may disturb adenylate cyclase system, which might be independent on cell growth. On the other hand, these PGs inhibited the incorporation of [3H]inositol into phospholipid fraction within 6 hrs. Furthermore, PGJ2 and Δ12PGJ2 inhibited carbachol- and/or histamine-induced accumulation of inositol phosphates with a similar dose-dependency to their inhibitions of cell growth. In membrane preparations, however, PGJ2 and Δ12PGJ2 failed to inhibit GTPγS (10 μM)- nor Ca2+ (1mM)-induced accumulation of inositol phosphate. The site of PGJ2 or Δ12PGJ2 in inhibition of inositol phosphate accumulation would not be phospholipase C nor a putative GTP binding protein involved in activation of phospholipase C. The present results indicate that PGJ2 and Δ12PGJ2 inhibit cell growth in human astrocytoma cells and the inhibition of phosphoinositide turnover by these PGs might be involved in the inhibition of cell growth. 相似文献
20.
de Gouvêa PF Soriani FM Malavazi I Savoldi M Goldman MH Loss O Bignell E da Silva Ferreira ME Goldman GH 《Fungal genetics and biology : FG & B》2008,45(7):1135-1146
Phosphate is an ion that is essential for fungal growth. The systems for inorganic phosphate (Pi) acquisition in eukaryotic cells (PHO) have been characterized as a low-affinity (that assures a supply of Pi at normal or high external Pi concentrations) and a high-affinity (activated in response to Pi starvation). Here, as an initial step to understand the PHO pathway in Aspergillus fumigatus, we characterized the PHO80 homologue, PhoBPHO80. We show that the ΔphoBPHO80 mutant has a polar growth defect (i.e., a delayed germ tube emergence) and, by phenotypic and phosphate uptake analyses, establish a link between PhoBPHO80, calcineurin and calcium metabolism. Microarray hybridizations carried out with RNA obtained from wild-type and ΔphoBPHO80 mutant cells identify Afu4g03610 (phoDPHO84), Afu7g06350 (phoEPHO89), Afu4g06020 (phoCPHO81), and Afu2g09040 (vacuolar transporter Vtc4) as more expressed both in the ΔphoBPHO80 mutant background and under phosphate-limiting conditions of 0.1 mM Pi. Epifluorescence microscopy revealed accumulation of poly-phosphate in ΔphoBPHO80 vacuoles, which was independent of extracellular phosphate concentration. Surprisingly, a phoDPHO84 deletion mutant is indistinguishable phenotypically from the corresponding wild-type strain. mRNA analyses suggest that protein kinase A absence supports the expression of PHO genes in A. fumigatus. Furthermore, ΔphoBPHO80 and ΔphoDPHO84 mutant are fully virulent in a murine low dose model for invasive aspergillosis. 相似文献