• Journal of Life Science
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• v.24 no.12
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• pp.1276-1283
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• 2014

Cell adhesion molecules determine the cell-cell binding and the interactions between cells and extracellular signals. Cell-cell junctional complexes, which maintain the structural integrity of tissues, consist of more than 50 proteins including multi-PDZ domain protein 1 (MUPP1). MUPP1 contains 13 postsynaptic density-95/disks large/zonula occludens-1 (PDZ) domains and serves a scaffolding function for transmembrane proteins and cytoskeletal proteins or signaling proteins, but the mechanism how MUPP1 links and stabilizes the juxtamembrane proteins has not yet been elucidated. We used the yeast two-hybrid system to identify proteins that interact with PDZ domains of MUPP1. We found an interaction between MUPP1 and cell adhesion molecule 1 (Cadm1, also known as SynCAM1, Necl-2, or TSLC1). Cadm1 bound to the second PDZ domain of MUPP1. The carboxyl (C)-terminal end of Cadm1 has a type II PDZ-association motif (-Y-F-I) which was essential for the interaction with MUPP1 in the yeast two-hybrid assay. MUPP1 also bound to the C-terminal cytoplasmic tail region of other Cadm family members (Cadm2, Cadm3, and Cadm4). In addition, these protein-protein interactions were observed in the glutathione S-transferase (GST) pull-down assay and by co-immunoprecipitation. Anti-MUPP1 antibody co-immunoprecipitated Cadm1 and Cadm4 with MUPP1 from mouse brain extracts. These results suggest that MUPP1 could mediate interaction between Cadms and cytoskeletal proteins.

• Journal of Life Science
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• v.26 no.3
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• pp.282-288
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• 2016

Protein-protein interactions regulate the subcellular localization and function of receptors, enzymes, and cytoskeletal proteins. Proteins containing the postsynaptic density-95/disks large/zonula occludens-1 (PDZ) domain have potential to act as scaffolding proteins and play a pivotal role in various processes, such as synaptic plasticity, neural guidance, and development, as well as in the pathophysiology of many diseases. Multi-PDZ domain protein 1 (MUPP1), which has 13 PDZ domains, has a scaffolding function in the clustering of surface receptors, organization of signaling complexes, and coordination of cytoskeletal dynamics. However, the cellular function of MUPP1 has not been fully elucidated. In the present study, a yeast two-hybrid system was used to identify proteins that interacted with the N-terminal PDZ domain of MUPP1. The results revealed an interaction between MUPP1 and Wdpcp (formerly known as Fritz). Wdpcp was identified as a planar cell polarity (PCP) effector, which is known to have a role in collective cell migration and cilia formation. Wdpcp bound to the PDZ1 domain but not to other PDZ domains of MUPP1. The C-terminal end of Wdpcp was essential for the interaction with MUPP1 in the yeast two-hybrid assay. This interaction was further confirmed in a glutathione S-transferase (GST) pull-down assay. When coexpressed in HEK-293T cells, Wdpcp was coimmunoprecipitated with MUPP1. In addition, MUPP1 colocalized with Wdpcp at the same subcellular region in cells. Collectively, these results suggest that the MUPP1-Wdpcp interaction could modulate actin cytoskeleton dynamics and polarized cell migration.

• Journal of Life Science
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• v.24 no.8
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• pp.820-826
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• 2014

The localization to specific subcellular sites and the regulation of cell surface receptors and channels are crucial for proper functioning. Postsynaptic density-95/Disks large/Zonula occludens-1 (PDZ)-domain is involved in recognition of and interaction between various proteins, by which the localization and the regulation are mediated. Multi-PDZ domain protein 1 (MUPP1) contains 13 PDZ domains. MUPP1 serves a scaffolding function for structure proteins and signaling proteins, but the mechanism how MUPP1 is stabilized and signalized has not yet been elucidated. We used the yeast two-hybrid system to identify proteins that interact with PDZ domains of MUPP1. We found an interaction between MUPP1 and Parkin. Parkin is an E3 ubiquitin ligase. Loss-of-function mutations of Parkin gene are known to cause an autosomal recessive juvenile parkinsonism. Parkin bound to the $12^{th}$ PDZ domain, but not to other PDZ domains of MUPP1. The C-terminal end of Parkin has a type II PDZ-association motif, which was essential for the interaction with MUPP1 in the yeast two-hybrid assay. When co-expressed in HEK-293T cells, Parkin co-localized with MUPP1. When co-expressed with ubiquitin in HEK-293T cells, MUPP1 has been strongly ubiquitinated by Parkin. These findings collectively suggest that MUPP1 is a novel substrate of Parkin and its function or stability could be modulated by Parkin-mediated ubiquitination.

• Bulletin of the Korean Chemical Society
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• v.33 no.5
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• pp.1707-1714
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• 2012

Grb2 is an adapter protein involved in the signal transduction and cell communication. The Grb2 is responsible for initiation of kinase signaling by Ras activation which leads to the modification in transcription. Ligand based pharmacophore approach was applied to built the suitable pharmacophore model for Grb2. The best pharmacophore model was selected based on the statistical values and then validated by Fischer's randomization method and test set. Hypo1 was selected as a best pharmacophore model based on its statistical values like high cost difference (182.22), lowest RMSD (1.273), and total cost (80.68). It contains four chemical features, one hydrogen bond acceptor (HBA), two hydrophobic (HY), and one ring aromatic (RA). Fischer's randomization results also shows that Hypo1 have a 95% significant level. The correlation coefficient of test set was 0.97 which was close to the training set value (0.94). Thus Hypo1 was used for virtual screening to find the potent inhibitors from various chemical databases. The screened compounds were filtered by Lipinski's rule of five, ADMET and subjected to molecular docking studies. Totally, 11 compounds were selected as a best potent leads from docking studies based on the consensus scoring function and critical interactions with the amino acids in Grb2 active site.

• Journal of Ginseng Research
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• v.44 no.3
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• pp.461-474
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• 2020

Background: Ginseng effectively reduces fatigue in both animal models and clinical trials. However, the mechanism of action is not completely understood, and its molecular targets remain largely unknown. Methods: By screening for proteins that interact with the primary components of ginseng (ginsenosides) in an affinity chromatography assay, we have identified muscle-type creatine kinase (CK-MM) as a potential target in skeletal muscle tissues. Results: Biolayer interferometry analysis showed that ginsenoside metabolites, instead of parent ginsenosides, had direct interaction with recombinant human CK-MM. Subsequently, 20(S)-protopanaxadiol (PPD), which is a ginsenoside metabolite and displayed the strongest interaction with CK-MM in the study, was selected as a representative to confirm direct binding and its biological importance. Biolayer interferometry kinetics analysis and isothermal titration calorimetry assay demonstrated that PPD specifically bound to human CK-MM. Moreover, the mutation of key amino acids predicted by molecular docking decreased the affinity between PPD and CK-MM. The direct binding activated CK-MM activity in vitro and in vivo, which increased the levels of tissue phosphocreatine and strengthened the function of the creatine kinase/phosphocreatine system in skeletal muscle, thus buffering cellular ATP, delaying exercise-induced lactate accumulation, and improving exercise performance in mice. Conclusion: Our results suggest a cellular target and an initiating molecular event by which ginseng reduces fatigue. All these findings indicate PPD as a small molecular activator of CK-MM, which can help in further developing better CK-MM activators based on the dammarane-type triterpenoid structure.

• Journal of Life Science
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• v.28 no.3
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• pp.284-292
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• 2018

The melanin-concentrating hormone (MCH), a cyclic hypothalamic peptide composed of 17 amino acids, was initially identified in chum salmon (Oncorhynchus keta) as a regulator of pigmentation. Mammalian MCHs are cyclic hypothalamic peptides composed of 19 amino acids that regulate food intake and energy homeostasis. The present study examined not only MCH expression of different tissues but also the melanohore aggregation and intracellular $Ca^{2+}$ influx of fMCH and the other MCH. Real-time qPCR showed that MCH expressed specially in the brain, gonad, and ovary, and expression of MCH was observed during the developmental stages. In the application of synthetic fMCH and both types of synthetic fMCH, dN-fMCH and dC-fMCH, scale melanophore induced significant changes in aggregation activity with various concentrations of MCH. Also, compared to hMCH and sMCH, fMCH exhibited a 36~99.85% increase in relative potency (%), whereas aggregation of dN-fMCH and dC-fMCH remained in a high concentration. However, dispersion was induced rapidly according to be low concentration of dN-fMCH and dC-fMCH. We show that fMCH and its derivates were bound human MCHR1 and rat MCHR expressed in HEK293T cells with nano-molar affinity and are likely to be ligand-induced to mobilize intracellular $Ca^{2+}$. These results may provide new ligands for binding assay with MCHew ligands, as a structure similar to the mammalian MCH structure was discovered in fish. Once the fMCH receptor system is in place, it can be compared to the MCH system of mammals in terms of MCH function.

• Korean Institute of Interior Design Journal
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• no.14
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• pp.82-89
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• 1998

During the boom of museum building in the 1970s-80s two museum wee erected in Germany. They are James Stirling's Neue Stattsgalerie Stuttgart and hans Hollein's Stadtisches Museum Abteiberg Monchengladbach, These two museums share the folowing point of similarity : Stirling's Neue Stattsgalerie Stuttgart and Hollein's Staditisches Museum Abetiberg Monchenbladbach both manifest the conditions of the times in light of their respective cities' local characters and historical contexts without being bound to traditional formality. Stirling and Hollein attempted to grasp the meaning of the 기nuseum as a city in miniature." Taking into account the t two museums’s territorial characteristics of being situated on a slope, both Stirling and Hollein made the walker to pass t through their building complex and introduced the concept of a public square within them. As a result, the museums are not j just two large buildings but are architecture composed of a collage of various structures. S Stirling’s architecture employs the method of attaching additional elements on top of basic constituents, which is suggestive '||'&'||'#61551; of the historical fragment from Shinkel's Altes Museum. On the other hand, Hollein applies a collage-style method as if he w were doing urban planning, maintaining the distinctiveness of each of the various forms and materials of buildings. T The object style buildings of the two museums actively demonstrate the contrast of double meaning to represent the a ambiguous and multifarious characteristics of the modern times. Stirling explores the theme of opposition and coexistence in h his Neue Stattsgalerie Stuttgart by placing a series of opposing concepts, such as the past and present and histor${\gamma}$ and t technology, in one space. Thereby, the contradiction and its appeal are manifested. Hollein made use of the visual illusion and c contradiction in alluding to the irony of the modern reality induced by nature and culture and history and technology. F For the above reasons and methods, James Stirling ’ s Neue Stattsgalerie Stuttgart allowed the penetration of art into daily l life and became the general public’s favored museum for its free and unrestrained environment. Likewise, Hans Hollein’s S Stadtisches Museum Abteiberg Moncbengladbach contribute to the development of museum function as it an expression of art i in the form of a complex piece of scupture on its own.ts own.

• Journal of the Korean Society of Combustion
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• v.18 no.4
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• pp.37-43
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• 2013

The effects of preferential diffusion of hydrogen in interacting counterflow $H_2$-air and CO-air premixed flames were investigated numerically. The global strain rate was varied in the range $30-5917s^{-1}$, where the upper bound of this range corresponds to the flame-stretch limit. Preferential diffusion of hydrogen was studied by comparing flame structures for a mixed average diffusivity with those where the diffusivities of H, $H_2$ and $N_2$ were assumed to be equal. Flame stability diagrams are presented, which show the mapping of the limits of the concentrations of $H_2$ and CO as a function of the strain rate. The main oxidation route for CO is $CO+O_2{\rightarrow}CO_2+O$, which is characterized by relatively slow chemical kinetics; however, a much faster route, namely $CO+OH{\rightarrow}CO_2+H$, can be significant, provided that hydrogen from the $H_2$-air flame is penetrated and then participates in the CO-oxidation. This modifies the flame characteristics in the downstream interaction between the $H_2$-air and CO-air flames, and can cause the interaction characteristics at the rich and lean extinction boundaries not to depend on the Lewis number of the deficient reactant, but rather to depend on chemical interaction between the two flames. Such anomalous behaviors include a partial opening of the upper lean extinction boundary in the interaction between a lean $H_2$-air flame and a lean CO-air flame, as well as the formation of two islands of flame sustainability in a partially premixed configuration with a rich $H_2$-air flame and a lean CO-air flame. At large strain rates, there are two islands where the flame can survive, depending on the nature of the interaction between the two flames. Furthermore, the preferential diffusion of hydrogen extends both the lean and the rich extinction boundaries.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.3
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• pp.398-407
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• 2020

Objective: The Yip1 domain family (YIPF) proteins were proposed to function in endoplasmic reticulum (ER) to Golgi transport and maintenance of the morphology of the Golgi, which were homologues of yeast Yip1p and Yif1p. YIPF3, the member 3 of YIPF family was a homolog of Yif1p. The aim of present study was to investigate the expression and regulation mechanism of porcine YIPF3. Methods: Quantitative realtime polymerase chain reaction (qPCR) was used to analyze porcine YIPF3 mRNA expression pattern in different tissues and pig kidney epithelial (PK15) cells stimulated by polyinosine-polycytidylic acid (poly [I:C]). Site-directed mutations combined with dual luciferase reporter assays and electrophoretic mobility shift assay (EMSA) were employed to reveal transcription regulation mechanism of porcine YIPF3. Results: Results showed that the mRNA of porcine YIPF3 (pYIPF3) was widely expressed with the highest levels in lymph and lung followed by spleen and liver, while weak in heart and skeletal muscle. Subcellular localization results indicated that it expressed in Golgi apparatus and plasma membranes. Upon stimulation with poly (I:C), the level of this gene was dramatically up-regulated in a time- and concentration-dependent manner. pYIPF3 core promoter region harbored three cis-acting elements which were bound by ETS proto-oncogene 2 (ETS2), zinc finger and BTB domain containing 4 (ZBTB4), and zinc finger and BTB domain containing 14 (ZBTB14), respectively. In which, ETS2 and ZBTB4 both promoted pYIPF3 transcription activity while ZBTB14 inhibited it, and these three transcription factors all played important regulation roles in tumorigenesis and apoptosis. Conclusion: The pYIPF3 mRNA expression was regulated by ETS2, ZBTB4, and ZBTB14, and its higher expression in immune organs might contribute to enhancing ER to Golgi transport of proteins, thus adapting to the immune response.

• Journal of the Korea Organic Resources Recycling Association
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• v.22 no.1
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• pp.27-34
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• 2014

demand for a low-cost treatment technology is high because the sewage sludge has an 80% or higher water content and a high energy consumption cost. This study apply the thermal hydrolysis reaction that consumes a small amount of energy for sludge treatment. The purpose of this study is to quantify the viscosity of sewage sludge according to reaction temperature. we measured continuously the torque of dewatered sludge by the reaction temperature. As the reaction temperature increased, the dewatered sludge is thermal hydrolysis under a high temperature and pressure. Therefore, the bond water in the sludge cells comes out as free water, which changes the dewatered sludge from a solid phase to slurry of a liquid phase. The results of the viscosity measurements according to the reaction temperature showed that the viscosity was very high at $270,180kg/m{\cdot}sec$ at a temperature of 293K, but rapidly decreased with increases in the reaction temperature to $12kg/m{\cdot}sec$ at a temperature of 400K and to $4kg/m{\cdot}sec$ at a temperature of 460K or higher, similar to the changes in the viscosity of water. And we was obtained the viscosity function of boundary condition for the optimal design of thermal hydrolysis reactor by numerical modeling based on the this results.