Iron-sulfur (Fe-S) clusters are one of the most ancient yet essential cofactors mediating various essential biological processes. In prokaryotes, Fe-S clusters are generated via several distinctive biogenesis mechanisms, among which the ISC (Iron-Sulfur Cluster) mechanism plays a house-keeping role to satisfy cellular needs for Fe-S clusters. The Escherichia coli ISC mechanism is maintained by several essential protein factors, whose structural characterization has been of great interest to reveal mechanistic details of the Fe-S cluster biogenesis mechanisms. In particular, nuclear magnetic resonance (NMR) spectroscopic approaches have contributed much to elucidate dynamic features not only in the structural states of the protein components but also in the interaction between them. The present minireview discusses recent advances in elucidating structural features of IscU, the key player in the E. coli ISC mechanism. IscU accommodates exceptional structural flexibility for its versatile activities, for which NMR spectroscopy was particularly successful. We expect that understanding to the structural diversity of IscU provides critical insight to appreciate functional versatility of the Fe-S cluster biogenesis mechanism.
Pseudomonas fluorescens GL20 is a plant growth-promoting rhizobacterium that produces a large amount of hydroxamate siderophore under iron-limited conditions. The strain GL20 considerably inhibited the spore germination and hyphal growth of a plant pathogenic fungus, Fusarium solani, when iron was limited, significantly suppressed the root-rot disease on beans caused by F. solani, and enhanced the plant growth. The mechanism for the beneficial effect of strain GL20 on the disease suppression was due to the siderophore production, evidenced by mutant strains derived from the strain. Analysis of the outer membrane protein profile revealed that the growth of strain GL20 induced the synthesis of specific iron-regulated outer membrane proteins with molecular masses of 85- and 90 kDa as the high-affinity receptors for the ferric siderophore. In addition, a cross-feeding assay revealed the presence of multiple inducible receptors for heterologous siderophores in the strain. In order to induce increased efficacy and potential in biological control of plant disease, a siderophore-overproducing mutant, GL20-S207, was prepared by NTG mutagenesis. The mutant GL20-S207 produced nearly 2.3 times more siderophore than the parent strain. In pot trials of beans with F. solani, the mutant increased plant growth up to 1.5 times compared with that of the parent strain. These results suggest that the plant growth-promoting P. fluorescens GL20 and the genetically bred P. fluorescens GL20-S207 can play an important role in the biological control of soil-borne plant diseases in the rhizosphere.
The objective of this study is to propose an alternative process for the small sewage treatment plants in rural communities. A biofilter has been used for biological wastewater treatment, which is becoming the alternative to the conventional activated sludge system. The proposed process used in this study, which is packed with floating media (i.e. expanded polystylene), has advantages of biofilter system and alternative flow system and they are incorporated into one process. Pilot and bench scale studies were performed using domestic wastewater. In the results of pilot plant study, it was observed that the stable effluent water quality was achieved and it met the present effluent criteria of suspended solid (SS), organic matters, T-N and T-P. In the study for determination of the cycle of backwashing, it was observed that the cycle of backwashing depended on BOD loading rates of influents. In the BOD loading rates of $0.5kg\;BOD/m^3{\cdot}day$ and $1.0kg\;BOD/m^3{\cdot}day$, the backwashing cycle of 28 hour and 16 hour were needed, respectively. The optimum backwashing time was 120~80 seconds at the media expansion rate of 50%. In the removal of SS, organic matters, T-N and T-P, SS removal was rather achieved by physical filtration than biological mechanism and the removal of organic matters except for SS, T-N and T-P were mainly rather achieved by biological mechanism than physical filtration. In bench-scale study, the effects of recirculation rate was investigated on removal of SS, TCOD, T-N and T-P. It was observed that the recirculation made removal efficiencies of SS, TCOD, T-N and T-P increased. Especially, in T-N removal, the increase of T-N removal efficiency of 40% was observed in the reicirculation rate of 1Q compared with 0Q.
Seo, Won-Sang;Oh, Han-Na;Park, Woo-Jung;Um, Sang-Young;Lee, Dae-Woo;Kang, Sang-Mo
KSBB Journal
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v.29
no.1
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pp.50-57
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2014
NF-${\kappa}B$ is a transcriptional factor which is involved in many biological processes including immunity, inflammation, and cell survival. Many investigators studied on the mechanism involved in activation of NF-${\kappa}B$ signalling pathway via ubiquitination and degradation of $I{\kappa}B$ regarding skin disease. Some specific molecules including Akt, MEK, p38 MAP Kinase, Stat3, et al. represent convergence points and key regulatory proteins in signaling pathways controlling cellular events such as growth and differentiation, energy homeostasis, and the response to stress and inflammation. Ultraviolet (UV) irradiation has many adverse effects on skin, including inflammation, alteration in the extracellular matrix, cellular senescence, apoptosis and skin cancer. Prunus mume, a naturally derived plant extract, has beneficial biological activities as blood fluidity improvement, anti-fatigue action, antioxidative and free radical scavenging activities, inhibiting the motility of Helicobacter pyolri. Previous reports on various beneficial function prompted us to investigate UVB-induced or other immunostimulated biological marker regarding P. mume extract. P. mume extract suppresses UVB-induced cyclooxygenase-2 (COX-2) expression in mouse skin epidermal JB6 P+ cells. The activation of activator protein-1 and nuclear factor-${\kappa}B$ induced by UVB was dose-dependently inhibited by P. mume extract treatment. This results suggest that P. mume extracts might be used as a potential agents for protection of inflammation or UVB induced skin damage.
Hur, Jae-Young;Kang, Gum-Yong;Choi, Min-Yeon;Jung, Jin Woo;Kim, Kwang-Pyo;Park, Sang-Hyun
Molecules and Cells
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v.26
no.1
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pp.41-47
/
2008
Mitogen-activated protein kinase (MAPK) signaling is a crucial component of eukaryotic cells; it plays an important role in responses to extracelluar stimuli and in the regulation of various cellular activities. The signaling cascade is evolutionarily conserved in the eukaryotic kingdom from yeast to human. In response to a variety of extracellular signals, MAPK activity is known to be regulated via phosphorylation of a conserved $T{\times}Y$ motif at the activation loop in which both threonine and tyrosine residues are phosphorylated by the upstream kinase. However, the mechanism by which both residues are phosphorylated continues to remain elusive. In the budding yeast, Saccharomyces cerevisiae, Fus3 MAPK is involved in the mating signaling pathway. In order to elucidate the functional mechanism of MAPK activation, we quantitatively profiled phosphorylation of the $T{\times}Y$ motif in Fus3 using mass spectrometry (MS). We used synthetic heavy stable isotope-labeled phosphopeptides and nonphosphopeptides corresponding to the proteolytic $T{\times}Y$ motif of Fus3 and accompanying data-dependent tandem MS to quantitatively monitor dynamic changes in the phosphorylation events of MAPK. Phosphospecific immunoblotting and the MS data suggested that the tyrosine residue is dynamically phosphorylated upon stimulation and that this leads to dual phosphorylation. In contrast, the magnitude of threonine phosphorylation did not change significantly. However, the absence of a threonine residue leads to hyperphosphorylation of the tyrosine residue in the unstimulated condition, suggesting that the threonine residue contributes to the control of signaling noise.
Translationally controlled tumor protein (TCTP), also known as histamine releasing factor (HRF), is found abundantly in different eukaryotic cell types. The sequence homology of TCTP between different species is very high, belonging to the MSS4/DSS4 superfamily of proteins. TCTP is involved in both cell growth and human late allergy reaction, as well as having a calcium binding property; however, its primary biological functions remain to be clearly elucidated. In regard to many possible functions, the TCTP of Plasmodium falciparum (Pf) is known to bind with an antimalarial agent, artemisinin, which is activated by heme. It is assumed that the endoperoxide-bridge of artemisinin is opened up by heme to form a free radical, which then eventually alkylates, probably to the Cys14 of PfTCTP. Study of the docking of artemisinin with heme, and subsequently with PfTCTP, was carried out to verify the above hypothesis on the basis of structural interactions. The three dimensional (3D) structure of PfTCTP was built by homology modeling, using the NMR structure of the TCTP of Schizosaccharomyces pombe as a template. The quality of the model was examined based on its secondary structure and biological function, as well as with the use of structure evaluating programs. The interactions between artemisinin, heme and PfTCTP were then studied using the docking program, FlexiDock. The center of the peroxide bond of artemisinin and the Fe of heme were docked within a short distance of $2.6{\AA}$, implying the strong possibility of an interaction between the two molecules, as proposed. When the activated form of artemisinin was docked on the PfTCTP, the C4-radical of the drug faced towards the sulfur of Cys14 within a distance of $2.48{\AA}$, again suggesting the possibility of alkylation having occurred. These results confirm the proposed mechanism of the antimalarial effect of artemisinin, which will provide a reliable method for establishing the mechanism of its biological activity using a molecular modeling study.
So, Soonku;Han, Kyeongsuk;Kim, Muyeol;Park, Hyerim;Seo, Eunkyoung;Kim, Yang-Pyo;Kim, Tae-Heung
Korean Journal of Plant Taxonomy
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v.38
no.1
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pp.43-50
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2008
The pollination system of Bupleurum latissimum Nakai (Apiaceae) was investigated in the natural population of Korea. The various insects of 19 species, 11 families, 5 orders visited the flowers of B. latissimum. Episyrphus balteatus and Lasioglossum occidens were considered as the most effective pollen vector which have associated specially with B. latissimum. The visitation frequency peaked at 10 AM - 13 PM and no visitor was recognized during night time. The flowers of B. latissimum last during only three days and they are protandry. It is also confirmed that the flower of B. latissimum is self-compatible and cross-pollination by vectors is critical for successful seed setting.
The objective of this study was to clarify the characteristics of the removed micropollutant since the breakthrough of adsorption ability was occurred in biological activated carbon(BAC) process. The removal efficiency of DOC (Dissolved Organic Carbon) was 36 % in the breakthrough of BAC occurred by NOM (Natural Organic Matter). The most of removal DOC was found out the adsorbable and biodegradable DOC (A&BDOC). But it was not clear to remove by any mechanism because A&BDOC have simultaneously the adsorption of activated carbon and biodegradation by microorganism in BAC. The removal of bromophenol was examined with BAC and rapid sand filter, for investigation of DOC removal mechanism in the breakthrough of BAC. In this experiment, BAC filter has been operated for 20 months for the treatment of reservoir water. The BAC filter was already exhausted by NOM. Bromophenol, adsorbable and refractory matter, was completely removed by BAC filter. Therefore, it might be removed by the adsorption in BAC. Adsorption isotherms of bromophenol were compared to two BACs which was preloaded with 500 daltons and 3,000 daltons of NOM. BAC preloaded with 3,000 daltons of NOM was not decreased to the adsorbability of bromophenol but BAC preloaded with 500 daltons of NOM was greatly decreased to it. These result indicated that NOM of low molecular weight can be removed by adsorption after a long period of operation and the breakthrough by NOM in BAC. Therefore, micropollutants might be removed through adsorption by saturated BAC.
Kim, Dong-Hak;Lim, Young-Ran;Park, Hyoung-Goo;Kim, Beom-Joon;Chun, Young-Jin
Toxicological Research
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v.25
no.1
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pp.35-40
/
2009
TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin) and related halogenated aromatic hydrocarbons elicit a diverse spectrum of biochemical and toxic responses in laboratory animals and mammalian cells in culture. Toxicity and carcinogenicity of TCDD is well established but the molecular mechanism is still poorly understood. Here, we found the noble responsive genes to TCDD using the differential display analysis. Treatment of HepG2 cells with TCDD showed a significantly different mRNA expression pattern from the untreated cells in differential display analysis. The differentially displayed bands were isolated and used as probes in dot blot and Northern blot analyses. Of thirty-five isolated differentially displayed bands, only two bands were confirmed as positive in dot blot and Northern blot analyses. The nucleotides sequences of these clones were analyzed and the search of Genebank database revealed that one clone is highly homologous with RanBP2 (Ras-related nuclear protein binding protein2; 92%) and the other is an unknown gene. RanBP2 is a nucleoporin with SUMO E3 ligase activity that functions in both nucleocytoplasmic transport and mitosis and its role as a novel tumor suppressor has been recently proposed. Thus, these results may suggest the clue elucidating the toxic mechanism of TCDD through RanBP2.
Lee, Yong Yook;Seo, Hwi Won;Kyung, Jong-Su;Hyun, Sun Hee;Han, Byung Cheol;Park, Songhee;So, Seung Ho;Lee, Seung Ho;Yi, Eugene C.
Journal of Ginseng Research
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v.43
no.4
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pp.666-675
/
2019
Background: Korean Red Ginseng (KRG) has been widely used as an herbal medicine to normalize and strengthen body functions. Although many researchers have focused on the biological effects of KRG, more studies on the action mechanism of red ginseng are still needed. Previously, we investigated the proteomic changes of the rat spleen while searching for molecular signatures and the action mechanism of KRG. The proteomic analysis revealed that differentially expressed proteins (DEPs) were involved in the increased immune response and phagocytosis. The aim of this study was to evaluate the biological activities of KRG, especially the immune-enhancing response of KRG. Methods: Rats were divided into 4 groups: 0 (control group), 500, 1000, and 2000 mg/kg administration of KRG powder for 6 weeks, respectively. Isobaric tags for relative and absolute quantitation was performed with Q-Exactive LC-MS/MS to compare associated proteins between the groups. The putative DEPs were identified by a current UniProt rat protein database search and by the Gene Ontology annotations. Results: The DEPs appear to increase the innate and acquired immunity as well as immune cell movement. These results suggest that KRG can stimulate immune responses. This analysis refined our targets of interest to include the potential functions of KRG. Furthermore, we validated the potential molecular targets of the functions, representatively LCN2, CRAMP, and HLA-DQB1, by Western blotting. Conclusion: These results may provide molecular signature candidates to elucidate the mechanisms of the immune response by KRG. Here, we demonstrate a strategy of tissue proteomics for the discovery of the molecular function of KRG.
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