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1.
Following binding to cognate ligand, estrogen receptor (ER) β interacts with specific responsive elements of the target genes
and recruits a host of nuclear proteins for hormone dependent gene regulation. However, it is poorly known which proteins
interact with ER β in mouse brain and whether their interaction and expression change with age. In this report, we have used
his-tag mouse ER β for interaction with nuclear proteins of cerebral cortex of young (6 ± 1 weeks), adult (25 ± 2 weeks),
and old (70 ± 5 weeks) female mice. We have identified estrogen receptor-associated protein (ERAP) 140 as one of the interacting
proteins and studied its interaction by pull down immunoblotting, far-Western blotting and immunoprecipitation, and expression
by western blotting. The data show that ERAP 140 interacts with ER β and its interaction decreases but its expression increases
with age in mouse cerebral cortex, suggesting its role in estrogen-mediated brain functions during aging. 相似文献
2.
Yu Yangsheng Bai Gang Liu Chunqin Li Yang Jin Yongjie Yang Wenbo 《Frontiers of Biology in China》2007,2(4):391-396
L-cysteine desulfhydrase (CD) plays an important role in L-cysteine decomposition. To identify the CD gene in Pseudomonas sp. TS1138 and investigate its effect on the L-cysteine biosynthetic pathway, the CD gene was cloned from Pseudomonas sp. TS1138 by polymerase chain reaction (PCR) method. The nucleotide sequence of CD gene was determined to be 1,215 bp, and
its homology with other sequences encoding CD was analyzed. Then the CD gene was subcloned into pET-21a(+) vector and expressed
in Escherichia coli (E. coli) by isopropyl-β-D-thiogalactopyranoside (IPTG) inducement. The recombinant CD was purified by Ni-NTA His-Bind resin, and its activity was
identified by the CD activity staining. The enzymatic properties of the recombinant CD were characterized and its critical
role involved in the L-cysteine biosynthetic pathway was also discussed.
__________
Translated from Microbiology, 2006, 33(4): 21–26 [译自: 微生物学通报] 相似文献
3.
Carlos A. Penno Stuart A. Morgan Anna Vuorinen Daniela Schuster Gareth G. Lavery Alex Odermatt 《Journal of lipid research》2013,54(10):2874-2883
11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) mediates glucocorticoid activation and is currently considered as therapeutic target to treat metabolic diseases; however, biomarkers to assess its activity in vivo are still lacking. Recent in vitro experiments suggested that human 11β-HSD1 metabolizes the secondary bile acid 7-oxolithocholic acid (7-oxoLCA) to chenodeoxycholic acid (CDCA) and minor amounts of ursodeoxycholic acid (UDCA). Here, we provide evidence from in vitro and in vivo studies for a major role of 11β-HSD1 in the oxidoreduction of 7-oxoLCA and compare its level and metabolism in several species. Hepatic microsomes from liver-specific 11β-HSD1-deficient mice were devoid of 7-oxoLCA oxidoreductase activity. Importantly, circulating and intrahepatic levels of 7-oxoLCA and its taurine conjugate were significantly elevated in mouse models of 11β-HSD1 deficiency. Moreover, comparative enzymology of 11β-HSD1-dependent oxidoreduction of 7-oxoLCA revealed that the guinea-pig enzyme is devoid of 7-oxoLCA oxidoreductase activity. Unlike in other species, 7-oxoLCA and its glycine conjugate are major bile acids in guinea-pigs. In conclusion, the oxidoreduction of 7-oxoLCA and its conjugated metabolites are catalyzed by 11β-HSD1, and the lack of this activity leads to the accumulation of these bile acids in guinea-pigs and 11β-HSD1-deficient mice. Thus, 7-oxoLCA and its conjugates may serve as biomarkers of impaired 11β-HSD1 activity. 相似文献
4.
Ghrelin is a hormone made in the stomach and known primarily for its growth hormone releasing and orexigenic properties. Nevertheless, ghrelin through its receptor, the GHS-R1a, has been shown to exert many roles including regulation of glucose homeostasis, memory & learning, food addiction and neuroprotection. Furthermore, ghrelin could promote overall health and longevity by acting directly in the immune system and promoting an extended antigen repertoire. The development of mice lacking either ghrelin (ghrelin−/−) or its receptor (ghsr−/−) have provided a valuable tool for determining the relevance of ghrelin and its receptor in these multiple and diverse roles. In this review, we summarize the most important findings and lessons learned from the ghrelin−/− and ghsr−/− mice. 相似文献
5.
Angelos Thanassoulas Michail Nomikos Maria Theodoridou Drakoulis Yannoukakos Dimitris Mastellos George Nounesis 《Biochimica et Biophysica Acta - Proteins and Proteomics》2010,1804(9):1908-1916
The BRCA1-associated RING domain protein 1 (BARD1) is the heterodimeric partner of BRCA1. The BRCA1/BARD1 complex demonstrates ubiquitin ligase activity and has been implicated in genomic stability and tumor suppression. Both proteins possess a structurally conserved C-terminal domain (BRCT). While BRCA1–BRCT has been shown to mediate BRCA1 interactions with phosphoproteins such as BRIP1 by recognizing the pSer-X-X-Phe motif, attempts to demonstrate analogous interactions of its dimeric counterpart BARD1–BRCT, have so far been unsuccessful. In this study, chemical-denaturation experiments of BARD1–BRCT domain suggest that its low thermodynamic stability (ΔG = 2.5 kcal/mol) at room temperature, may affect some of its biochemical properties, such as its interaction with phosphopeptides. The stability of BARD1–BRCT domain at 10 °C, increases to 7.5 kcal/mol and isothermal titration calorimetry (ITC) experiments at this lower temperature showed binding to the BRIP1 phosphopeptide via an enthalpy-driven interaction, which appears to be specific to the pSer-X-X-Phe peptide-binding motif. Substitution of either pSer at position 0 with Ser (non-phosphorylated peptide) or Phe with Val at position + 3, leads to no-binding ITC results. While these findings are indicative that BRIP1 is a potential BARD1 binding partner, it becomes evident that in vitro binding assays involving the entire BARD1 protein and in vivo experiments are also needed to establish its binding partners and its potential role in tumor suppression pathways. 相似文献
6.
Streptococcus pneumoniae is a common bacterial pathogen that is well known for its ability to cause acute respiratory disease (pneumonia), ear infections, and other serious illnesses. This Gram-positive bacterium relies on its carbohydrate-metabolizing capabilities for full virulence in its host; however, the range of glycan targets that it can attack is presently not fully appreciated. S. pneumoniae is known to have a fucose utilization operon that in the TIGR4 strain plays a role in its virulence. Here we identify a second type of fucose utilization operon that is present in a subset of S. pneumoniae strains, including the serotype 3 strain SP3-BS71. This operon contains a transporter with a solute-binding protein, FcsSBP (fucose solute-binding protein), that interacts tightly (Ka ∼ 1 × 106 M− 1) and specifically with soluble A- and B-antigen trisaccharides but displays no selectivity between these two sugars. The structure of the FcsSBP in complex with the A-trisaccharide antigen, determined to 2.35 Å, reveals its mode of binding to the reducing end of this sugar, thus highlighting this protein's requirement for soluble blood group antigen ligands. Overall, this report exposes a heretofore unknown capability of certain S. pneumoniae strains to transport and potentially metabolize the histo-blood group antigen carbohydrates of its host. 相似文献
7.
Qin Li Xin Guan Karen Yen Jiyuan Zhang Jiusheng Yan 《The Journal of general physiology》2016,147(4):337-351
The large-conductance, calcium-activated potassium (BK) channels consist of the pore-forming, voltage- and Ca2+-sensing α subunits (BKα) and the tissue-specific auxiliary β and γ subunits. The BK channel γ1 subunit is a leucine-rich repeat (LRR)–containing membrane protein that potently facilitates BK channel activation in many tissues and cell types through a vast shift in the voltage dependence of channel activation by ∼140 mV in the hyperpolarizing direction. In this study, we found that the single transmembrane (TM) segment together with its flanking charged residues is sufficient to fully modulate BK channels upon its transplantation into the structurally unrelated β1 subunit. We identified Phe273 and its neighboring residues in the middle of the TM segment and a minimum of three intracellular juxtamembrane Arg residues as important for the γ1 subunit’s modulatory function and observed functional coupling between residues of these two locations. We concluded that the TM segment is a key molecular determinant for channel association and modulation and that the intracellular positively charged cluster is involved mainly in channel association, likely through its TM-anchoring effect. Our findings provide insights into the structure–function relationship of the γ1 subunit in understanding its potent modulatory effects on BK channels. 相似文献
8.
The new ligand 4′-(4?-pyridyl-N-oxide)-2,2′:6′,2″-terpyridine (pyNoxterpy) and its homoleptic iron(II) complex have been synthesised, and structural and spectroscopic studies have been carried out. The obtained results have been compared with the reported data for the parent ligand 4′-(4?-pyridyl)-2,2′:6′,2″-terpyridine (pyterpy) and its homoleptic iron(II) complex. Significant differences between the spectral and electrochemical properties of the metal complexes have been found, derived from the changes in the electronic properties of the coordinated ligands. 相似文献
9.
Tamoxifen is extensively metabolized, and several metabolites have been detected in human serum. The aim of this study was to examine the interaction of human serum albumin (HSA) with tamoxifen and its metabolites 4-hydroxytamoxifen and endoxifen at physiological conditions, using constant protein concentration and various drug contents. FTIR, UV-Visible, CD and fluorescence spectroscopic methods as well as molecular modeling were used to analyse drug binding mode, the binding constant and the effects of drug complexation on HSA stability and conformation. Structural analysis showed that tamoxifen and its metabolites bound HSA via both hydrophobic and hydrophilic interactions with overall binding constants of Ktam = 1.8 (±0.2) × 104 M−1, K4-hydroxytam = 1.8 (±0.4) × 104 M−1 and Kendox = 2.0 (±0.5) × 104 M−1. The number of bound drugs per protein is 1.2 (tamoxifen), 1.7 (4-hydroxitamoxifen) and 1.0 (endoxifen). Structural modeling showed the participation of several amino acid residues in drug-HSA complexation, with extended H-bonding network. HSA conformation was altered by tamoxifen and its metabolites with a major reduction of α-helix and an increase in β-sheet, random coil and turn structures, indicating a partial protein unfolding. Our results suggest that serum albumins can act as carrier proteins for tamoxifen and its metabolites in delivering them to target tissues. 相似文献
10.
The serine/threonine kinase polo-like kinase 1 (Plk1) is critically involved in multiple mitotic processes and has been established as an adverse prognostic marker for tumor patients. Plk1 localizes to its substrates and its intracellular anchoring sites via its polo-box domain (PBD), which is unique to the family of polo-like kinases. Therefore, inhibition of the Plk1 PBD has been suggested as an approach to the inhibition of Plk1 that circumvents specificity problems associated with the inhibition of the conserved adenosine triphosphate (ATP) binding pocket. Here we report on the development of a high-throughput assay based on fluorescence polarization that allows the discovery of small-molecule inhibitors of the Plk1 PBD. The assay is based on binding of the Plk1 PBD to a phosphothreonine-containing peptide comprising its optimal binding motif with a Kd of 26 ± 2 nM. It is stable with regard to dimethyl sulfoxide (DMSO) and time, and it has a Z′ value of 0.73 ± 0.06 in a 384-well format. 相似文献
11.
The paradox high zooplankton biomass-low vegetal particulate organic matter in high turbidity zones: What way for energy transfer? 总被引:1,自引:0,他引:1
Valérie David Benoît Sautour Pierre Chardy 《Journal of experimental marine biology and ecology》2006,333(2):202-218
Estuarine ecosystems are characterized by high zooplanktonic biomasses, essentially constituted by copepods and mysids whose nutritional requirements are mainly provided by phytoplankton, an easily available carbon form. The Gironde estuary is characterized by high turbidities which limit light penetration in the water column and therefore primary production. Consequently, primary production is low and its availability for higher trophic level is very limited. The main goal of this study was to characterize the total vegetal particulate organic matter (POM) in high turbidity zones of the Gironde estuary during summer (a critical period characterized by high heterotrophic bacterial degradation and high zooplanktonic biomasses) and to analyse its utilization by zooplankton, using prey/predator experiments and trophic biomarkers (fatty acids). The specific goals were to define (i) how vegetal POM was exploited by the different zooplanktonic groups (protozoa, copepods and mysids) and (ii) which alternative preys could be used when vegetal POM was not sufficient to ensure their nutritional requirements.Chlorophyll biomass was very low in the MTZ during summer 2002 (0.48 ± 0.03 mg m− 3). Total zooplankton grazing was low (19% d− 1) probably due to a large contribution of detritus originating from terrestrial plants in vegetal POM compared to phytoplankton. The highest grazing pressure was exercised by the mysid Mesopodopsis slabberi due to its high abundances and by its almost entirely herbivorous diet (phytoplankton and small terrestrial detritus). Grazing rates (19.7 ± 4.2 and 9.6 μgC cop− 1 d− 1 for juveniles and adults, respectively) seemed to be sufficient to satisfy their daily carbon requirement. Grazing rate of the copepod Eurytemora affinis (139 ngC cop− 1 d− 1) seemed to be insufficient to cover its nutritional requirements and the copepods probably needed to complete a great part of their diet from protozoa. Grazing rates of the mysid Neomysis integer (24.7 ± 0.01 and 20.89 ± 8.45 μgC cop− 1 d− 1 for juveniles and adults, respectively) were higher than those of M. slabberi when feeding only on phytoplankton. However, when other preys were introduced in its environment, N. integer only fed on the copepod E. affinis with a preference for nauplii. The study revealed the great importance of protozoa and bacteria in the trophic transfers between vegetal POM and zooplankton in the MTZ during summer, despite the low protozoa grazing pressure on vegetal POM (3.1%). The detritic food chain probably implies various trophic transfers with little direct relationships between vegetal POM and zooplankton. 相似文献
12.
Biodegradation of glyphosate in sod-podzol soil by both the indigenous micro flora and the introduced strain Ochrobactrum anthropi GPK 3 was studied with respect to its sorption and mobility. The experiments were carried out in columns simulating the vertical
soil profile. Soil samples studied were taken from soil horizons 0–10, 10–20, and 20–30 cm deep. It was found out that the
most of the herbicide (up to 84%) was adsorbed by soil during the first 24 h; the rest (16%) remained in the soluble fraction.
The adsorbed glyphosate was completely extractable by alkali. No irreversible binding of glyphosate was observed. By the end
of the experiment (21st day), glyphosate was only found in extractable fractions. The comparison of the effect of the introduced
O. anthropi GPK 3 and indigenous microbial community on the total toxicant content (both soluble and absorbed) in the upper 10 cm soil
layer showed its reduction by 42% (21 mg/kg soil) and 10–12% (5 mg/kg soil), respectively. Simultaneously, 14–18% glyphosate
moved to a lower 10–20 cm layer. Watering (that simulated rainfall) resulted in a 20% increase of its content at this depth;
6–8% of herbicide was further washed down to the 20–30 cm layer. The glyphosate mobility down the soil profile reduced its
density in the upper layer, where it was available for biodegradation, and resulted in its concentration in lower horizons
characterized by the absence (or low level) of biodegradative processes. It was shown for the first time how the herbicide
biodegradation in soil can be increased manifold by introduction of the selected strain O. anthropi GPK 3. 相似文献
13.
14.
Peifeng Hou Ching-Ying Kuo Chun-Ting Cheng Jing-Ping Liou David K. Ann Qiang Chen 《PloS one》2014,9(11)
Hypoxia-inducible factor 1α (HIF-1α), a major mediator of tumor physiology, is activated during tumor progression, and its abundance is correlated with therapeutic resistance in a broad range of solid tumors. The accumulation of HIF-1α is mainly caused by hypoxia or through the mutated succinate dehydrogenase A (SDHA) or fumarate hydratase (FH) expression to inhibit its degradation. However, its activation under normoxic conditions, termed pseudohypoxia, in cells without mutated SDHA or FH is not well documented. Here, we show that dimethyl-2-ketoglutarate (DKG), a cell membrane-permeable precursor of a key metabolic intermediate, α-ketoglutarate (α-KG), known for its ability to rescue glutamine deficiency, transiently stabilized HIF-1α by inhibiting activity of the HIF prolyl hydroxylase domain-containing protein, PHD2. Consequently, prolonged DKG-treatment under normoxia elevated HIF-1α abundance and up-regulated the expression of its downstream target genes, thereby inducing a pseudohypoxic condition. This HIF-1α stabilization phenotype is similar to that from treatment of cells with desferrioxamine (DFO), an iron chelator, or dimethyloxalyglycine (DMOG), an established PHD inhibitor, but was not recapitulated with other α-KG analogues, such as Octyl-2KG, MPTOM001 and MPTOM002. Our study is the first example of an α-KG precursor to increase HIF-1α abundance and activity. We propose that DKG acts as a potent HIF-1α activator, highlighting the potential use of DKG to investigate the contribution of PHD2-HIF-1α pathway to tumor biology. 相似文献
15.
Elekes K Sandor K Moricz A Kereskai L Kemeny A Szoke E Perkecz A Reglodi D Hashimoto H Pinter E Szolcsanyi J Helyes Z 《Peptides》2011,32(7):1439-1446
The presence of pituitary adenylate cyclase-activating polypeptide (PACAP) and its receptors in capsaicin-sensitive peptidergic sensory nerves, inflammatory and immune cells suggest its involvement in inflammation. However, data on its role in different inflammatory processes are contradictory and there is little known about its functions in the airways. Therefore, our aim was to examine intranasal endotoxin-induced subacute airway inflammation in PACAP gene-deficient (PACAP−/−) and wild-type (PACAP+/+) mice. Airway responsiveness to inhaled carbachol was determined in unrestrained mice with whole body plethysmography 6 h and 24 h after LPS. Myeloperoxidase (MPO) activity referring to the number of accumulated neutrophils and macrophages was measured with spectrophotometry and interleukin-1β (IL-1β) concentration with ELISA from the lung homogenates. Histological evaluation and semiquantitative scoring were also performed. Bronchial responsiveness, as well as IL-1β concentration and MPO activity markedly increased at both timepoints. Perivascular edema dominated the histological picture at 6 h, while remarkable peribronchial granulocyte accumulation, macrophage infiltration and goblet cell hyperplasia were seen at 24 h. In PACAP−/− mice, airway hyperreactivity was significantly higher 24 h after LPS and inflammatory histopathological changes were more severe at both timepoints. MPO increase was almost double in PACAP−/− mice compared to the wild-types at 6 h. In contrast, there was no difference between the IL-1β concentrations of the PACAP+/+ and PACAP−/− mice. These results provide evidence for a protective role for PACAP in endotoxin-induced airway inflammation and hyperreactivity. 相似文献
16.
N. M. Jansonius J. H. van Hateren 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》1993,172(4):467-471
We recorded from the spiking on-off unit in the first optic chiasm (between lamina and medulla) in the blowfly Calliphora vicina, and investigated its spatial properties. The receptive field extends over (11.4±0.9)° horizontally and (8.7±0.6)° vertically, i.e. about 7 by 5 interommatidial angles. The line spread function of the on-off unit — calculated from its response to moving sinusoidal gratings — has a half-width of (2.3±0.2)°. This half-width is slightly broader than that of the photoreceptor. Lateral inhibition occurs when two different areas of the receptive field are stimulated simultaneously. Fast temporal adaptation (i.e. adaptation to trains of short light pulses) takes place independently in different areas of the receptive field. 相似文献
17.
Surgucheva I Park BC Yue BY Tomarev S Surguchov A 《Cellular and molecular neurobiology》2005,25(6):1009-1033
Summary Mutations in the gene encoding human myocilin are associated with some cases of juvenile and early-onset glaucoma. Glaucomatous
mutations prevent myocilin from being secreted. The analysis of the defects associated with mutations point to the existence
of factor(s) in addition to mutations that might be implicated in the development of glaucoma. In the present paper, we found
that interaction of myocilin with one of the members of the synuclein family alters its properties, including its ability
to be secreted. Results of immunoprecipitation show that myocilin is a γ-synuclein-interacting protein. Further analysis demonstrated
that both myocilin and γ-synuclein are expressed in human TM cells, immortalized rat ganglion (RGC-5) cells, and HT22 hippocampal
neurons. According to Western blotting, in addition to monomeric form with molecular weight 17 kDa γ-synuclein is present
as higher molecular weight forms (∼35 and 68 KDa), presumably dimer and tetramer. Myocilin and γ-synuclein have partially
overlapping perinuclear localization. Dexamethasone upregulates myocilin expression in RGC-5 cells and HT22 hippocampal neurons.
We found alterations of myocilin properties as a result of its interaction with γ-synuclein. In cultured cells, γ-synuclein
upregulates myocilin expression, inhibits its secretion and prevents the formation of high molecular weight forms of myocilin.
Although both α-synuclein and γ-synuclein are expressed in HTM cells, only γ-synuclein interacts with myocilin and alters
its properties.
We conclude that myocilin and γ-synuclein interact and as a result, myocilin's properties are changed. Since myocilin and
γ-synuclein have partially overlapping intracellular localization in cell types that are implicated in glaucoma development,
their interaction may play an important role in glaucoma. 相似文献
18.
The new species Polhillia ignota Boatwr. is described. This species is known only from two collections, one between Vredenburg and Saldanha and the other close to Porterville. The new species is most similar to P. obsoleta, which is known only from a few localities around Worcester, in its narrow, sericeous leaflets and flowers of equal size, but differs in its flattened mature leaves with larger leaflets (up to ± 12 mm long), longer pseudo-peduncles (up to ± 15 mm long), denser inflorescences (with up to four flowers), shorter pedicels (1–2 mm long) and non-auriculate wing- and keel petals. 相似文献
19.
Christine Elizabeth Cooper Ariovaldo P. Cruz-Neto 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》2009,179(6):773-781
The honey possum is the only non-volant mammal to feed exclusively on a diet of nectar and pollen. Like other mammalian and
avian nectarivores, previous studies indicated that the honey possum’s basal metabolic rate was higher than predicted for
a marsupial of equivalent body mass. However, these early measurements have been questioned. We re-examined the basal metabolic
rate (2.52 ± 0.222 ml O2 g−1 h−1) of the honey possum and confirm that it is indeed higher (162%) than predicted for other marsupials both before and after
accounting for phylogenetic history. This, together with its small body mass (5.4 ± 0.14 g; 1.3% of that predicted by phylogeny)
may be attributed to its nectarivorous diet and mesic distribution. Its high-basal metabolic rate is associated with a high-standard
body temperature (36.6 ± 0.48°C) and oxygen extraction (19.4%), but interestingly the honey possum has a high point of relative
water economy (17.0°C) and its standard evaporative water loss (4.33 ± 0.394 mg H2O g−1 h−1) is not elevated above that of other marsupials, despite its mesic habitat and high dietary water intake. 相似文献