• Korean Journal of Microbiology
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• v.40 no.2
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• pp.87-93
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• 2004

Dynamic modification of cytoplasmic and nuclear proteins by O-linked N-acetylglucosamine (O-GlcNAc) on Ser and Thr residues is ubiquitous in higher eukaryotes. And this modification may serve as a signaling mod-ification analogous to protein phosphorylation. Addition and cleavage of O-GlcNAc are catalyzed by O-linked GlcNAc transferase (OGT) and O-linked N-acety1glucosaminidase (O-GlcNAcase), respectively. Two types of human O-GlcNAcase gene were cloned and expressed as three fusion proteins in Escherichia coli. O-GlcNA-case activity showed in the order of thioredoxin fusion> $6{\times}His$ tag> GST fusion. O-GlcNAcase had enzy-matic activity against only ${\rho}$NP-GlcNAc of seven tested substrate analogs. Blast search revealed that O-GlcNAcase has two conserved domains, amino terminal hyaluronidase-like domain and carboxy terminal N-acetyltransferase domain. Extensive deletion studies were done to define catalytically important domains. The deletions of hyaluronidase-like domain and N-acetyltransferase domain abolished enzyme activity. But, N-ter-minal 55 amino acid deletion and C-terminal truncation showed lower activity. Based on deletion analysis, we suggest that hyaluronidase-like domain is essential for enzyme activity and carboxy terminal N-acetyltrans-ferase domain may be modulatory function.

• (The)Study of the Eastern Classic
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• no.66
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• pp.35-73
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• 2017

The aim of this article is to analyze the feature of the $Hongch^{\prime}{\breve{o}}n$ poetry society(紅泉社). The leader of the $Hongch^{\prime}{\breve{o}}n$ poetry society was Yi Mansu(李晩秀), and the members of that were Yi Naksu(李洛秀), Sin Chin(申縉) Sin Chak(申綽) Sin $Hy{\breve{o}}n$(申絢) Pak Chongu(朴宗羽) $Ch{\breve{o}}ng$ Sukwi(鄭遂龜) $Kw{\breve{o}}n$ Sik(權?) Kim Kyeon(金啓溫). They organized the poetry society at Gi-dae(企臺) of Seoul. The poems of the $Hongch^{\prime}{\breve{o}}n$ poetry society were included in "$K{\breve{u}}gw{\breve{o}}n$ yugo(?園遺稿)", a collection of Yi Mansu's works. The $Hongch^{\prime}{\breve{o}}n$ poetry society was maintained for three years, from 1817 to 1820. The members of the $Hongch^{\prime}{\breve{o}}n$ poetry society gathered at Kidae(企臺) and wrote poems together, such as the linked verses(聯句), divisions of rime(分 韻), replying rhyming verses(次韻) and so on. This article especially analyzed the linked verses of the $Hongch^{\prime}{\breve{o}}n$ poetry society. The following is a summary of characteristics in the linked verses of the $Hongch^{\prime}{\breve{o}}n$ poetry society. First, the members of the $Hongch^{\prime}{\breve{o}}n$ poetry society showed their poetic genius by writing the linked verses. They competitively designed unique words and techniques to exhibit their poetic genius. Especially, their poetic genius were exposed in $y{\breve{o}}n^{\prime}gu$(放雲樓聯句)>, modeling <$S{\breve{o}}ngnam$ $y{\breve{o}}n^{\prime}gu$(城南聯句)>'s style. The members of the $Hongch^{\prime}{\breve{o}}n$ poetry society had remarkable literary attainments. Second, the members of the $Hongch^{\prime}{\breve{o}}n$ poetry society promoted friendship by writing the linked verses. They expressed the pleasure of having a poetry party in the linked verses. Their linked verses are elegance. In addition, their poems are full of the pride as officials. And they were glad that they lived in the happy era. So they extoled the king's virtue in their linked verses.

• BMB Reports
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• v.29 no.3
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• pp.266-271
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• 1996

A recombinant human erythropoietin (EPO), expressed in Chinese hamster ovary (CHO) cells, is glycosylated at Asn 24, Asn 38, Asn 83, and Ser 126. After release of the N-linked carbohydrate chains by $peptide-N^{4}-(N-acetyl-{\beta}-glucosaminyl)$ asparagine amidase F, the oligosaccharides were analyzed by FACE (Fluorophore-Assisted Carbohydrate Electrophoresis). The O-linked carbohydrate chain was separated by hydrazine, and analyzed by FACE. The monosacccharide composition of recombinant EPO showed man nose, fucose, galactose, N-acetylglucosamine, N-acetylneuraminic acid, and a trace of N-acetylgalactosamine, which are typical monosaccharides in the glycoproteins from the CHO cell. Sequences of N-linked and O-linked oligosaccharides were determined. The structure and composition of oligosaccharides attached to recombinant human EPO, expressed in the CHO cell, are identical to the reported oligosaccharide structure in human EPO isolated from urine.

• Archives of Pharmacal Research
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• v.17 no.5
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• pp.309-313
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• 1994

Phytase of rat intestine had a large amount of oilgosacchanrides ; The enzyme consisted of two different subunits with the molecular weights of 90 KDa and 70 KDa in its intack form, whereas the apparent molecular weights tumed to 72 KDa and 52 KDa, respectively, after deglycosylation. The treatment with glycopeptidase F reduced the molecular weights from 90 KDa and 70 KDa to 83 KDa and 52 KDa, respectively, While endoglycosidase H caused no change in their molecular weights. These results indicate that the 70KDa subunit contains only the N-linked oilgosaccharide chains, while the 90KDa subunit ocntains O-linked oilgosaccharides as well as N-linked ones. Enzyme-linked lectin assays suggeted that bisecting N-acetyl-D-glucosamine and galactose 1-4 N-acetyl-D-glucosamine structures were present and that fucose was included in these oilgosaccharide moieties. Sialic acid was not found in either subunit.

• Analytical Science and Technology
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• v.17 no.1
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• pp.16-22
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• 2004

Inclusion complexes of the p-nitrophenol with ${\beta}$-cyclodextrin (CD), sulfated ${\beta}$-CD, and ${\beta}$-CD polymer cross-linked with epichlorohydrine (EP) were prepared and characterized by Raman spectroscopy. The intensity of vibration peaks of the C-O and C-N at 1284 and $856cm^{-1}$ of the p-nitrophenol in the presence of EP-linked CD polymer was remarkably increased, respectively. The vibration modes at 1284 and $856cm^{-1}$ are assigned to the out-of phase C-C-O stretching mode and the C-N stretching mode, respectively. The vibration peaks at 1284 and $856cm^{-1}$ increased with increasing the content of EP-linked CD polymers, while decreased with increasing the p-nitrophenol content. Furthermore, the vibration mode of the $NO_2$ symmetric stretch at $1344cm^{-1}$ enhanced with increasing the content of p-nitrophenol.

• Journal of Microbiology and Biotechnology
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• v.12 no.2
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• pp.306-311
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• 2002

O-linked N-acetylglucosamine (O-GlcNAc) is an abundant posttranslationally modified compound in eukaryotic cells. Human O-GlcNAc transferase (OGT) was produced as a maltose binding protein (MBP) fusion protein, which showed significant catalytic activity to modify recombinant Sp1, transcription factor. To facilitate the production of O-GlcNAc modified proteins, instead of using the tedious in vitro glycosylation reaction or expression in eukaryotic cells, a MBP-fusion OGT expression vector (pACYC184-MBPOGT) was constructed using pACYC184 plasmid, which could coexist with general prokaryotic expression vectors containing ColE1 origin. By cotransforming pACYC184-MBPOGT and pGEX-2T vectors into Escherichia coli BL21, intracellular O- GlcNAcylated proteins could be obtained by a simple purification procedure. It is expected that this may be a useful tool for production of O-GlcNAc modified proteins.

• Journal of the Korean Wood Science and Technology
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• v.42 no.6
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• pp.740-746
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• 2014

The unripe fruit of cudrania tricuspidata was extracted with 50% ethanol. The crude water-soluble extracts were separated by liquid-liquid separation with n-hexane, ethyl acetate and butanol followed by precipitation with ethanol, and then the water-soluble polysaccharide (F1) was isolated by the fractionation through gel permeation chromatography using preparative PLaquagel-OH column with water. The structure was characterized by monosaccharide composition with HPAEC-PAD, methylation analysis with GC-MS, FT-IR and HPLC. According to the data, F1 was com posed of glucose (22.84 mM), galactose (13.75 mM), arabinose (45.87 mM), xylose (7.49 mM). It was revealed which uronic acid and acetyl group were not attached in F1. And it is constituted of 1-linked arabinose, 1,4-linked arabinose, 1,3-linked glucose, 1,4-linked galactose, 1,4-linked glucose, 1,3,6-linked galactose, 1,3,6-linked glucose and the ratio was showed 1.1 : 1.0 : 4.9 : 7.5 : 3.0 : 3.1 : 1.4 : 1.5.

• Bulletin of the Korean Chemical Society
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• v.33 no.10
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• pp.3355-3360
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• 2012

To promote the photoelectric conversion efficiency of dye-sensitized solar cells (DSSCs), graphene is introduced as a working electrode with $TiO_2$ in this study, because it has great transparency and very good conductivity. XRD patterns indicate the presence of graphene and $TiO_2$ particles in graphene-linked $TiO_2$ samples. Moreover, TEM pictures also show that the nano-sized $TiO_2$ particles are highly dispersed and well-linked onto the thin layered graphene. On the basis of the UV-visible spectra, the band gaps of $TiO_2$, 1.0 wt % graphene-$TiO_2$, 5.0 wt % graphene-$TiO_2$, and 10.0 wt % graphene-$TiO_2$ are 3.16, 2.94, 2.25, and 2.11 eV, respectively. Compared to pure $TiO_2$, the energy conversion efficiency was enhanced considerably by the application of graphene-linked $TiO_2$ anode films in the DSSCs to approximately 6.05% for 0.1 wt % graphene-$TiO_2$ with N719 dye (10.0 mm film thickness and $5.0mm{\times}5.0mm$ cell area) under $100mW/cm^2$ of simulated sunlight. The quantum efficiency was the highest when 1.0 wt % of graphene was used. In impedance curves, the resistance was smallest for 1.0 wt % graphene-$TiO_2$-DSSC.

• Journal of Life Science
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• v.16 no.6
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• pp.955-959
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• 2006

A dynamic cycle of addition and removal of O-linked N-acetylglucosamine (O-GlcNAc) at serine and threonine residues is emerging as a key regulator of nuclear and cytoplasmic protein activity. In this work, immunocytochemistry was carried out to investigate the subcellular expression of GlcNAc kinase (NAGK, EC 2.7.1.59) that catalyzes the phosphorylation of GlcNAc to GlcNAc 6-phosphate. Immunostainings of cultured rat hippocampal neurons revealed patchy or punctate distribution of NAGK. When NAGK is doublestained with caveolin-1 or flotillin, markers for caveolae and lipid rafts, respectively, NAGK was co-localized with these markers. These results indicate that most, if not all, of the NAGK immunopunctae represent caveolae and lipid rafts, and suggest NAGK's role in these membrane microdomains.

• Korean Journal of Microbiology
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• v.40 no.2
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• pp.147-153
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• 2004

Protein modification by N-acetyl-${\beta}$-D-glucosamine (O-G1cNAc) on the hydroxyl groups of Ser or Thr ubiq-uitously occurs in eukaryotic cells and is involved in many cellular phenomena. The level of O-G1cNAc-mod-ified protein is regulated by OGT and O-GlcNAcase enzymes. We have tried to produce recombinant O-GlcNAcase in E. coli as an effort to establish in vitro screening system for modulators of O-GlcNAcase. The culture conditions for improvement of O-GlcNAcase productivity, were as follows: induction temperature, $30^{\circ}C$; the concentration of L-arabinose, 0.02% and induction time, 5 hr. Under these culture conditions, E. coli cells containing O-GlcNAcase gene had no enzyme activity until up to 3 hr culture. However, O-GlcNAcase activity dramatically increased from 3 to 5 hr culture. It almost maintained the same level after 5 hr culture. Western blot analysis verified the amount of expressed O-GlcNAcase increased with culture time, being con-sistent with activity data. The optimal reaction condition determined in this study was as follows: protein quan-tity, $5{\mu}g$; reaction time, 30 min; reaction temperature, $45^{\circ}C$; substrate concentration, 2 mM; reaction pH, 6.5. Methanol had little effect on O-GlcNAcase activity and 90% of activity were retained at 10%. Only 15% resid-ual activity were detected at 5% of chloroform.