• Molecules and Cells
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• v.37 no.4
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• pp.322-329
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• 2014

N-acetylglucosamine kinase (GlcNAc kinase or NAGK; EC 2.7.1.59) is highly expressed and plays a critical role in the development of dendrites in brain neurons. In this study, the authors conducted structure-function analysis to verify the previously proposed 3D model structure of GlcNAc/ATP-bound NAGK. Three point NAGK mutants with different substrate binding capacities and reaction velocities were produced. Wild-type (WT) NAGK showed strong substrate preference for GlcNAc. Conversion of Cys143, which does not make direct hydrogen bonds with GlcNAc, to Ser (i.e., C143S) had the least affect on the enzymatic activity of NAGK. Conversion of Asn36, which plays a role in domain closure by making a hydrogen bond with GlcNAc, to Ala (i.e., N36A) mildly reduced NAGK enzyme activity. Conversion of Asp107, which makes hydrogen bonds with GlcNAc and would act as a proton acceptor during nucleophilic attack on the ${\gamma}$-phosphate of ATP, to Ala (i.e., D107A), caused a total loss in enzyme activity. The overexpression of EGFP-tagged WT or any of the mutant NAGKs in rat hippocampal neurons (DIV 5-9) increased dendritic architectural complexity. Finally, the overexpression of the small, but not of the large, domain of NAGK resulted in dendrite degeneration. Our data show the effect of structure on the functional aspects of NAGK, and in particular, that the small domain of NAGK, and not its NAGK kinase activity, plays a critical role in the upregulation of dendritogenesis.

• Journal of Nutrition and Health
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• v.7 no.4
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• pp.12-20
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• 1974

Quantitative analysis was achieved by gas-liquid chromatographic method (GLC) with a single column system of OV-17 for 16 of protein amino acids in mushroom (agaricus bisporus). The quantities of protein amino acids in mushroom were determined $48.32{\sim}255.94mg%$ alanine, $108.6F{\sim}364.82mg%$ glycine, $124.30{\sim}314.17mg%$ Valine, $32.99{\sim}418.79mg%$ leucine and isoleucine, $151.78{\sim}669.07mg%$ threonine, $88.12{\sim}4F6.3Fmg%$ Serine, $21.9F{\sim}114.94mg%$ Hydroxyproline, $20.F4{\sim}174.63mg%$ proline, $34.52{\sim}173.59mg%$ Methionine, $225.25{\sim}1417.61mg%$ Aspartic Acid, $10F.00{\sim}392.17mg%$ Phenylalanine, $12F.46{\sim}535.65mg%$ Glutamic Acid and Lysine, Tyrosine in trace amount.

• BMB Reports
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• v.30 no.2
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• pp.95-100
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• 1997

Two kinds of cDNA encoding mouse $Gal{\beta}$1,3(4)GlcNAc ${\alpha}$2,3-sialyltransferase (mST3Gal III) and $Gal{\beta}$1,4(3)GlcNAc ${\alpha}$2,3-sialyltransferase (mST3Gal IV) were isolated from mouse brain cDNA library by means of a PCR-based approach. The cDNA sequences included an open reading frame coding for proteins of 374 and 333 amino acids, respectively, and the primary structure of these enzymes suggested a putative domain structure consisting of four regions, like that in other glycosyltransferases. The deduced amino acid sequences of mST3GaI III and IV showed a 98% and 89% identity with rat ST3GaI III and human ST3Gal IV, respectively. Northern analysis indicated that the expression of mST3Gal III mRNA was abundant in heart, liver and adult brain, while that of mST3GaI IV mRNA was detected in all tissues tested except for testis, but the level was the highest in liver. Soluble forms of mST3GaI III and IV transiently expressed in COS cells exhibited enzyme activity toward acceptor substrates containing the terminal either $Gal{\beta}$1,3GlcNAc or $Gal{\beta}$1,4GlcNAc sequences. The substrate preferences of both enzymes were stronger for tetrasaccharides than for disaccharides.

• Analytical Science and Technology
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• v.9 no.2
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• pp.123-133
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• 1996

The optimum conditions for the analysis of the difenoconazole fungicide on soil and crops were investigated and the residues of that in apple and soil were identified by using the gas chromatography. The extract with acetonitrile was separated with saturated NaCl and n-hexane solution after filtered, and concentrated. Obtained fungicide residues were transfered to the florisil column and eluted with acetone and n-hexane mixed solution for the analysis by GLC(ECD). From the standard addition experiments with 0.20 and 1.0ppm, the average recoveries were 86~92% and the detection limit was 0.01 ppm. It seems to be safely used when difenoconazole is treated three times until 15 days before harvest of apple. In this case residual amounts of difenoconazole in apple was from 0.037ppm to 0.044ppm. The soil samples extracted with methanol and ammonium hydroxide mixed solution were partitioned with dichloromethane and saturated sodium chloride solution. The organic phase was concentrated and redissolved with toluene and analyzed with GLC(FID) after cleaned with Sep-Pak column. From the standard addition experiments with 0.10, 0.50 and 1.0ppm, the average recoveries were 101.2~103.7% and the detection limit was 0.025ppm.

• Korean journal of applied entomology
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• v.13 no.4 s.21
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• pp.235-243
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• 1974

Recenty, the pesticides Pollutions in connection with the maintenance of dietary life are poised as serious sanitary problems. Author have established an appropriate analysis method in this connection and, at the same time, analysed the levels of pesticides residues, such as organochloride pesticides and organic mercurials, contained in main cereals (rice, barley, and wheat) collected throughout the country. Using gas liquid chromatography method, comparative analyses were made of organochloride Pesticides with electron capture detector, and organic mercurials with electron capture detector and the Dithizone Method. As a result, the organic mercurials analysis using gas liquid chromatography is believed to hold out an especially good method for both in terms of its sensitivity and its Practical applications. The summary obtained from these results is as follows; The detectable limit of organochloride pesticides is $5\times10^{-10}$ grams and that of organic mecurials is $5\times10^{-9}$ grams. The detections using the Dithizone Method are difficult. The gas liquid chromatographic analysis of organic mercurials is very simple in its operation and high in its sensitivity, compare with the analysis using the Dithizone Method. Therefore, this analysis is expected to be a good method applicable to the pesticide residues analysis. The levels of pesticide residues contained in samples are very little for tolerance and, therefore, no problem i.: foreseen for eating. The appropriate conditions of gas liquid chromatographic analysis obtained from these results are expected very useful for the date establishing an officially authorized analysis method.

• The Korean Journal of Pesticide Science
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• v.9 no.4
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• pp.311-315
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• 2005

The simultaneous or consecutive product analysis is needed for quality control because the various items are produced in pesticide manufacturer. This study was conducted to establish a multi-pesticide analysis making possible to analyze several active ingredients with one injection of mixed active ingredients under same instrument-condition in the cause of quality control with accuracy and speed. The test was conducted with 3 pesticides, iprobenfos 17%GR, isoprothiolane 12%GR, tebufenozide 20%SC and performed by GLC and HPLC. With the GLC method, 2 active ingredients of iprobenfos and isoprothiolane were analyzed but tebufenozide was not detective simultaneously. With the HPLC method, all of the active ingredients in those three pesticides were simultaneously analyzed in this study.

• Proceedings of the KIEE Conference
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• 2015.07a
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• pp.268-269
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• 2015

복잡화된 국내 전력계통의 부하는 지속적으로 증가하는 반면 새로운 설비의 건설이 어렵고, 지역 편중화된 발전설비 때문에 선로 과부하, 고장전류, 전압안정도 문제가 발생하고 있다. 초고압 선로의 고장은 계통을 크게 불안정하게 하기 때문에 고장에 의해 영향을 받는 지역과 고장 후 계통의 조류변화를 분석하는 것은 중요하다. 현재 고장의 영향을 분석하기 위하여 조류계산을 통한 정적해석과 시모의를 통한 동적해석을 사용하다. 그리고 좀 더 큰 그림을 그리기 위하여 각종 전압안정도 지수를 사용한다. 하지만 일반적으로는 고장이후 계통에서 유효전력 흐름에 변화가 있는 지역을 분석하기 위해서는 번거로운 작업이 필요한 단점이 있다. Generation loass coefficient(GLC)는 transmmision loss factor(TLF)에서 발생한 문제를 분석하기 위해 제안되었고, load loss coefficient(LLC)는 각 부하에 전력을 공급하기 위해 발생하는 손실을 발전기별로 분석하기 위해 제안되었다. 위의 두 지수는 계통해석을 위해서 제안된 것은 아니었으나 전력조류추적기법을 기반으로하여 개발되었기 때문에 계통의 전력조류 흐름 변화에 대한 정보를 담고 있다는 특징이 있다. 본 논문에서는 GLC와 LLC의 개념에 대하여 설명하고 계통에서 발생하는 고장의 영향을 해석하는 관점에서 GLC와 LLC를 활용한다. 시뮬레이션 결과를 통해 GLC와 LLC지수로 계통에 대한 이해를 높이는 방안에 대하여 제안한다.

• Journal of mucopolysaccharidosis and rare diseases
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• v.2 no.1
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• pp.13-16
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• 2016

Mucolipidosis (ML) II/III are autosomal recessive diseases caused by deficiency of post-translational modification of lysosomal enzymes. The mannose-6-phosphate (M6P) residue in lysosomal enzymes synthesized by N-acetylglucosamine 1-phosphotransferase (GlcNAc-phosphotransferase) serves as recognition marker for trafficking in lysosomes. GlcNAc-phosphotransferase is encoded by GNPTAB and GNPTG. Mutations in GNPTAB cause severe ML II alpha/beta and the attenuated ML III alpha/beta. Whereas mutations in GNPTG cause the ML III gamma, the attenuated type of ML III variant. For the diagnostic approaches, increased urinary oligosaccharides excretion could be a screening test in clinically suspicious patients. To confirm the diagnosis, instead of measuring the activity of GlcNAc phosphotransferase, measuring the enzymatic activities of different lysosomal hydrolases are useful for diagnosis. The activities of several lysosomal hydrolases are decreased in fibroblasts but increased in serum of the patients. In addition, the sequence analysis of causative gene is warranted. Therefore, the confirmatory diagnosis requires a combination of clinical evaluation, biochemical and molecular genetic testing. ML II/III show complex disease manifestations with lysosomal storage as the prime cellular defect that initiates consequential organic dysfunctions. As there are no specific therapy for ML to date, understanding the molecular pathogenesis can contribute to develop new therapeutic approaches ultimately.

• Bulletin of the Korean Chemical Society
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• v.30 no.11
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• pp.2723-2728
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• 2009

The conformational properties of oligosaccharides are important to understand carbohydrate-protein interactions. A trimannoside, methyl 3,6-di-O-($\alpha$-D-Man)-$\alpha$-D-Man (TRIMAN) is a basic unit of N-linked oligosaccharides. This TRIMAN moiety was further modified by GlcNAc (BISECT), which is important to biological activity of N-glycan. To characterize the trimannoside and its bisecting one we performed a molecular dynamics simulation in water. The resulting models show the conformational transition with two major and minor conformations. The major conformational transition results from the $\omega$ angle transition; another minor transition is due to the $\psi$ angle transition of $\alpha$ (1 $\rightarrow$ 6) linkage. The introduction of bisecting GlcNAc on TRIMAN made the different population of the major and minor conformations of the TRIMAN moiety. Omega ($\omega$) angle distribution is largely changed and the population of gt conformation is increased in BISECT oligosaccharide. The inter-residue hydrogen bonds and water bridges via bisecting GlcNAc residue make alterations on the local and overall conformation of TRIMAN moiety. These changes of conformational distribution for TRIMAN moiety can affect the overall conformation of N-glycan and the biological activity of glycoprotein.

• Molecules and Cells
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• v.24 no.2
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• pp.294-300
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• 2007

The mammalian glycosylphosphatidylinositol (GPI) anchor consists of three mannoses attached to acylated GlcN-(acyl)PI to form $Man_3$-GlcN-(acyl)PI. The first of the three mannose groups is attached to an intermediate to generate Man-GlcN-(acyl)PI by the first mannosyltransferase (GPI-MT-I). Mammalian and protozoan GPI-MT-I have different substrate specificities. PIG-M encodes the mammalial GPI-MT-I which has 423 amino acids and multiple transmembrane domains. In this work we cloned PIG-M homologues from humans, Plasmodium falciparum (PfPIG-M), and Saccharomyces cerevisiae (GPI14), to test whether they could complement GPI-MT-I-deficient mammalian cells, since this biosynthetic step is likely to be a good target for selective screening of inhibitors against many pathogenic organisms. PfPIG-M partially restored cell surface expression of the GPI-anchored protein CD59 in PIG-M deficient mammalian cells, and first mannose transfer activity in vitro; however, this was not the case for GPI14.