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1.
Key Amino Acid Residues Involved in Multi-Point Binding Interactions between Brazzein, a Sweet Protein, and the T1R2-T1R3 Human Sweet Receptor 总被引:1,自引:0,他引:1
Fariba M. Assadi-Porter Emeline L. Maillet James T. Radek John L. Markley 《Journal of molecular biology》2010,398(4):584-599
The sweet protein brazzein [recombinant protein with sequence identical with the native protein lacking the N-terminal pyroglutamate (the numbering system used has Asp2 as the N-terminal residue)] activates the human sweet receptor, a heterodimeric G-protein-coupled receptor composed of subunits Taste type 1 Receptor 2 (T1R2) and Taste type 1 Receptor 3 (T1R3). In order to elucidate the key amino acid(s) responsible for this interaction, we mutated residues in brazzein and each of the two subunits of the receptor. The effects of brazzein mutations were assayed by a human taste panel and by an in vitro assay involving receptor subunits expressed recombinantly in human embryonic kidney cells; the effects of the receptor mutations were assayed by in vitro assay. We mutated surface residues of brazzein at three putative interaction sites: site 1 (Loop43), site 2 (N- and C-termini and adjacent Glu36, Loop33), and site 3 (Loop9-19). Basic residues in site 1 and acidic residues in site 2 were essential for positive responses from each assay. Mutation of Y39A (site 1) greatly reduced positive responses. A bulky side chain at position 54 (site 2), rather than a side chain with hydrogen-bonding potential, was required for positive responses, as was the presence of the native disulfide bond in Loop9-19 (site 3). Results from mutagenesis and chimeras of the receptor indicated that brazzein interacts with both T1R2 and T1R3 and that the Venus flytrap module of T1R2 is important for brazzein agonism. With one exception, all mutations of receptor residues at putative interaction sites predicted by wedge models failed to yield the expected decrease in brazzein response. The exception, hT1R2 (human T1R2 subunit of the sweet receptor):R217A/hT1R3 (human T1R3 subunit of the sweet receptor), which contained a substitution in lobe 2 at the interface between the two subunits, exhibited a small selective decrease in brazzein activity. However, because the mutation was found to increase the positive cooperativity of binding by multiple ligands proposed to bind both T1R subunits (brazzein, monellin, and sucralose) but not those that bind to a single subunit (neotame and cyclamate), we suggest that this site is involved in subunit-subunit interaction rather than in direct brazzein binding. Results from this study support a multi-point interaction between brazzein and the sweet receptor by some mechanism other than the proposed wedge models. 相似文献
2.
Mehdi Adib Fariba Peytam Reihaneh Shourgeshty Maryam Mohammadi-Khanaposhtani Mehdi Jahani Somaye Imanparast Mohammad Ali Faramarzi Bagher Larijani Ali Akbar Moghadamnia Ensieh Nasli Esfahani Fatemeh Bandarian Mohammad Mahdavi 《Bioorganic & medicinal chemistry letters》2019,29(5):713-718
Twenty three fused carbazole–imidazoles 6a–w were designed, synthesized, and screened as new α-glucosidase inhibitors. All the synthesized fused carbazole-imidazoles 6a-w were found to be more active than acarbose (IC50?=?750.0?±?1.5?µM) against yeast α-glucosidase with IC50 values in the range of 74.0?±?0.7–298.3?±?0.9?µM. Kinetic study of the most potent compound 6v demonstrated that this compound is a competitive inhibitor for α-glucosidase (Ki value?=?75?µM). Furthermore, the in silico studies of the most potent compounds 6v and 6o confirmed that these compounds interacted with the key residues in the active site of α-glucosidase. 相似文献
3.
Evidence for Growth of Enterococci in Municipal Oxidation Ponds, Obtained Using Antibiotic Resistance Analysis
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Elaine Moriarty Fariba Nourozi Beth Robson David Wood Brent Gilpin 《Applied microbiology》2008,74(23):7204-7210
The Christchurch wastewater treatment plant uses a series of six oxidation ponds to reduce the bacterial load of treated effluent before it is discharged into the local estuary. To ensure that this discharge does not adversely affect water quality in the receiving environment, local regulations specify maximum levels in the discharge for a number of parameters, including enterococci. Between 2001 and 2006, regulations required fewer than 300 enterococci per 100 ml in summer. During this period, the discharge intermittently exceeded this limit, with unexplained levels of enterococci of up to 180,000/100 ml. Characterization of these enterococci by antibiotic resistance analysis showed that enterococci sampled over 4 months had almost identical resistance profiles. In contrast, enterococci from raw sewage and wildfowl from around the oxidation ponds had a diverse range of antibiotic resistance profiles that could be distinguished from each other and also from those of enterococci from the discharge. The hypothesis of a clonal nature of the enterococci in the discharge was supported by molecular genotype analysis, suggesting that these bacteria may have replicated in the pond environment rather than being reflective of breakthrough in the sewage treatment process or the result of recent wildfowl inputs to the ponds. This study highlights the usefulness of antibiotic resistance analysis in identifying this phenomenon and is the first report of apparent replication of a specific type of enterococci in an oxidation pond environment. 相似文献
4.
Gholamhossein Sodeifian Fariba Razmimanesh 《Journal of biomolecular structure & dynamics》2019,37(7):1666-1684
In this research, for the first time, molecular dynamics (MD) method was used to simulate aspirin and ibuprofen at various concentrations and in neutral and charged states. Effects of the concentration (dosage), charge state, and existence of an integral protein in the membrane on the diffusion rate of drug molecules into lipid bilayer membrane were investigated on 11 systems, for which the parameters indicating diffusion rate and those affecting the rate were evaluated. Considering the diffusion rate, a suitable score was assigned to each system, based on which, analysis of variance (ANOVA) was performed. By calculating the effect size of the indicative parameters and total scores, an optimum system with the highest diffusion rate was determined. Consequently, diffusion rate controlling parameters were obtained: the drug–water hydrogen bond in protein-free systems and protein–drug hydrogen bond in the systems containing protein. 相似文献
5.
Amirhosein NaghshNilchi Laleh Ebrahimi Ghahnavieh Fariba Dehghanian 《Journal of cellular and molecular medicine》2022,26(16):4530
Platinum resistance is one of the major concerns in ovarian cancer treatment. Recent evidence shows the critical role of epithelial–mesenchymal transition (EMT) in this resistance. Epithelial‐like ovarian cancer cells show decreased sensitivity to cisplatin after cisplatin treatment. Our study prospected the association between epithelial phenotype and response to cisplatin in ovarian cancer. Microarray dataset was acquired from the GEO database. After identifying differentially expressed genes (DEGs) between epithelial‐like and mesenchymal‐like cells, the module identification analysis was performed using weighted gene co‐expression network analysis (WGCNA). The gene ontology (GO) and pathway analyses of the most considerable modules were performed. The protein–protein interaction network was also constructed. The hub genes were specified using Cytoscape plugins MCODE and cytoHubba, followed by the survival analysis and data validation. Finally, the co‐expression of miRNA‐lncRNA‐TF with the hub genes was reconstructed. The co‐expression network analysis suggests 20 modules relating to the Epithelial phenotype. The antiquewhite4, brown and darkmagenta modules are the most significant non‐preserved modules in the Epithelial phenotype and contain the most differentially expressed genes. GO, and KEGG pathway enrichment analyses on these modules divulge that these genes were primarily enriched in the focal adhesion, DNA replication pathways and stress response processes. ROC curve and overall survival rate analysis show that the co‐expression pattern of the brown module''s hub genes could be a potential prognostic biomarker for ovarian cancer cisplatin resistance. GSE47856相似文献
6.
Intraguild predation by the generalist predator Orius majusculus on the parasitoid Encarsia formosa 总被引:1,自引:0,他引:1
Fariba Sohrabi Annie Enkegaard Parviz Shishehbor Moosa Saber Mohammad Said Mosaddegh 《BioControl》2013,58(1):65-72
Intraguild predation of Orius majusculus (Reuter) (Heteroptera: Anthocoridae) on Encarsia formosa (Gahan) (Hymenoptera: Aphelinidae), both natural enemies of Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae), was studied under laboratory conditions. The experiments quantified prey consumption by 5th instar nymphs and adults of O. majusculus offered unparasitised 3rd, early 4th or 4th instar B. tabaci nymphs or parasitised nymphs containing 2nd or 3rd larval instar or pupal parasitoids. In addition, prey preference of the two stages of O. majusculus for parasitised or unparasitised whitefly nymphs was studied using nine different prey combinations. Both predator stages readily preyed upon on both unparasitised and parasitised B. tabaci. In no-choice experiments, predation on 3rd instar E. formosa by adult predators was the highest, while predator nymphs preyed most on unparasitised 3rd instar B. tabaci and 2nd instar parasitoids. Predation of predator stages was lowest on 4th instar B. tabaci and E. formosa pupae. In all prey combinations, both stages of O. majusculus showed a significant preference for parasitised over unparasitised whitefly nymphs except for the combination of 5th instars of O. majusculus with early 4th instar whiteflies and E. formosa pupae. The results indicate that intraguild interactions between O. majusculus and E. formosa may have negative effects on biological control of B. tabaci. 相似文献
7.
Neural responses to visual stimuli are strongest in the classical receptive field, but they are also modulated by stimuli in a much wider region. In the primary visual cortex, physiological data and models suggest that such contextual modulation is mediated by recurrent interactions between cortical areas. Outside the primary visual cortex, imaging data has shown qualitatively similar interactions. However, whether the mechanisms underlying these effects are similar in different areas has remained unclear. Here, we found that the blood oxygenation level dependent (BOLD) signal spreads over considerable cortical distances in the primary visual cortex, further than the classical receptive field. This indicates that the synaptic activity induced by a given stimulus occurs in a surprisingly extensive network. Correspondingly, we found suppressive and facilitative interactions far from the maximum retinotopic response. Next, we characterized the relationship between contextual modulation and correlation between two spatial activation patterns. Regardless of the functional area or retinotopic eccentricity, higher correlation between the center and surround response patterns was associated with stronger suppressive interaction. In individual voxels, suppressive interaction was predominant when the center and surround stimuli produced BOLD signals with the same sign. Facilitative interaction dominated in the voxels with opposite BOLD signal signs. Our data was in unison with recently published cortical decorrelation model, and was validated against alternative models, separately in different eccentricities and functional areas. Our study provides evidence that spatial interactions among neural populations involve decorrelation of macroscopic neural activation patterns, and suggests that the basic design of the cerebral cortex houses a robust decorrelation mechanism for afferent synaptic input. 相似文献
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9.
Fariba Mehrkhou Mehdi Taheri Sarhozaki 《Archives Of Phytopathology And Plant Protection》2013,46(12):1444-1453
Effect of four different cole crops (Brassica oleracea var. botrytis, Brassica oleracea var. capitata, Brassica oleracea var. italica and Brassica oleracea var. viridis) on biological parameters of the large white butterfly Pieris brassicae was evaluated at temperature 26 ± 1 °C, 60 ± 5% R. H. and a photoperiod of 16: 8 (L:D) h. The shortest larval and pupal period stages were recorded on B. oleracea var. botrytis (22.18 ± 0.20 days) and (13.32 ± 0.17 days), respectively. The life span was longest on B. oleracea var. viridis (60.43 ± 2.34 days) and shortest on B. oleracea var. botrytis (50.19 ± 0.51 days). The highest percentage of larval and pupal mortality was observed on B. oleracea var. viridis (74%), and (53%), respectively. We found that P. brassicae prefers B. oleracea var. botrytis and B. oleracea var. capitata among cole crops; it is due to the lowest percentage of larval and pupal mortality and the highest rate of life table parameters, including survival rate (lx) and life expectancy (ex), which makes them to be susceptible varieties to this pest. Contrary, a longer developmental time on B. oleracea var. viridis may be attributed to poor nutritional status and reduced survival of the cohort, resulting in high rates of mortality, which was partial resistance to pest. Knowledge of the biology and life table parameters of P. brassicae on different cole crops could be effective in detecting and monitoring the pest infestation, variety selection and crop breeding. 相似文献
10.
Claudia C. Cornilescu Gabriel Cornilescu Hongyu Rao Sarah F. Porter Marco Tonelli Michele L. DeRider John L. Markley Fariba M. Assadi‐Porter 《Proteins》2013,81(6):919-925
The sweet protein brazzein, a member of the Csβα fold family, contains four disulfide bonds that lend a high degree of thermal and pH stability to its structure. Nevertheless, a variable temperature study has revealed that the protein undergoes a local, reversible conformational change between 37 and 3°C with a midpoint about 27°C that changes the orientations and side‐chain hydrogen bond partners of Tyr8 and Tyr11. To test the functional significance of this effect, we used NMR saturation transfer to investigate the interaction between brazzein and the amino terminal domain of the sweet receptor subunit T1R2; the results showed a stronger interaction at 7°C than at 37°C. Thus the low temperature conformation, which alters the orientations of two loops known to be critical for the sweetness of brazzein, may represent the bound state of brazzein in the complex with the human sweet receptor. Proteins 2013; © 2012 Wiley Periodicals, Inc. 相似文献