• Journal of Life Science
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• v.19 no.8
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• pp.1062-1066
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• 2009

N-Acetylglucosamine kinase (GlcNAc kinase or NAGK; EC 2.7.1.59) catalyzes the phosphorylation of GlcNAc to GlcNAc-6-phosphate (GlcNAc-6-P). Despite detailed characterization of the enzyme itself, there have been few studies on the expression of NAGK in mammalian tissues. In the rat hippocampal neuron in culture, NAGK-immunoreactivity (IR) formed clusters in somatodendritic domains. In this study we characterized the NAGK clusters that protrude out the long axis of dendritic shafts. By double-labeling of the neurons with antibodies against NAGK and various synaptic proteins, we show that NAGK is positioned at the base of spines, while there were no NAGK protrusions into inhibitory postsynaptic sites. Immunoblot analysis showed that NAGK was included in synaptosomes but not in PSD fractions. Our results indicate that the NAGK clusters at the dendritic periphery protrude into spines.

• 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.

• Molecules and Cells
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• v.40 no.7
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• pp.476-484
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• 2017

C-X-C chemokine receptor 4 (CXCR4) stimulates cancer metastasis. NF-${\kappa}B$ regulates CXCR4 expression in cancer cells, and O-GlcNAc modification of NF-${\kappa}B$ promotes its transcriptional activity. Here, we determined whether CXCR4 expression is affected by O-GlcNAcylation of NF-${\kappa}B$ in lung metastasis of cervical cancer. We found elevated levels of O-linked-N-actylglucosamine transferase (OGT) and O-GlcNAcylation in cervical cancer cells compared to those in non-malignant epithelial cells and detected increased expression of NF-${\kappa}B$ p65 (p65) and CXCR4 in cervical cancer cells. Knockdown of OGT inhibited the O-GlcNAcylation of p65 and decreased CXCR4 expression levels in HeLa cells. Thiamet G treatment increased O-GlcNAcylated p65, which subsequently enhanced CXCR4 expression levels. Inhibition of O-GlcNAcylation by 6-Diazo-5-oxo-L-norleucine (DON) treatment decreased p65 activation, eventually inhibiting CXCR4 expression in HeLa cells. Lung tissues from mice engrafted with OGT-knockdown HeLa cells (shOGT) exhibited lower expression of Ki-67 and HPV E6 and E7 oncogenes compared to lung tissues from mice engrafted with control HeLa cells (shCTL). In addition, lung tissues from mice engrafted with shOGT cells exhibited lower p65 and CXCR4 immunoreactivity compared to tissues from mice engrafted with shCTL cells. Taken together, our data suggest that p65 O-GlcNAcylation promotes lung metastasis of cervical cancer cells by activating CXCR4 expression.

• Journal of Food Hygiene and Safety
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• v.3 no.4
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• pp.203-209
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• 1988

This study was performed to develop the simple and rapid determination method of preservatives in Yogurt and Soybean.sauce. 1. The analytical method by HPLC system was as follow: The sample was diluted, centrifuged, filtered, if necessary, and analyzed by HPLC system with UV detector. 2. The analytical method by GLC system was as follow: The sample was extracted directly with ether, added cone. sulfuric acid to destroy emulsion and analyzed by the GLC system with FID. 3. The recovery rates of preservatives by the above methods were higher than 99.0 %. 4. Total running time for the above methods was less than 50 minutes. Especially, the running time for dilution method by HPLC system was one-third of that for GLC method.

• Journal of Nutrition and Health
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• v.48 no.4
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• pp.319-326
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• 2015

Purpose: Borage oil (BO) and safflower oil (SO) are efficacious in reversing epidermal hyperproliferation, which is caused by the disruption of epidermal barrier. In this study, we compared the antiproliferative effect of dietary BO and SO. Altered metabolism of ceramide (Cer), the major lipid of epidermal barrier, was further determined by measurement of epidermal levels of individual Cer, glucosylceramide (GlcCer), and sphingomyelin (SM) species, and protein expression of Cer metabolizing enzymes. Methods: Epidermal hyperproliferation was induced in guinea pigs by a hydrogenated coconut diet (HCO) for 8 weeks. Subsequently, animals were fed diets of either BO (group HCO + BO) or SO (group HCO + SO) for 2 weeks. As controls, animals were fed BO (group BO) or HCO (group HCO) diets for 10 weeks. Results: Epidermal hyperproliferation was reversed in groups HCO + BO (67.6% of group HCO) and HCO + SO (84.5% of group HCO). Epidermal levels of Cer1/2, GlcCer-A/B, and ${\beta}$-glucocerebrosidase (GCase), an enzyme of GlcCer hydrolysis for Cer generation, were higher in group HCO + BO than in group HCO, and increased to levels similar to those of group BO. In addition, epidermal levels of SM1, serine palmitoyltransferase (SPT), and acidic sphingomyelinase (aSMase), enzymes of de novo Cer synthesis and SM hydrolysis for Cer generation, but not of Cer3-7, were higher in group HCO + BO than in group HCO. Despite an increase of SPT and aSMase in group HCO + SO to levels higher than in group HCO, epidermal levels of Cer1-7, GlcCer-A/B, and GCase were similar in these two groups. Notably, acidic ceramidase, an enzyme of Cer degradation, was highly expressed in group HCO + SO. Epidermal levels of GlcCer-C/D and SM-2/3 did not differ among groups. Conclusion: Dietary BO was more prominent for reversing epidermal hyperproliferation by enhancing Cer metabolism with increased levels of Cer1/2, GlcCer-A/B, and SM1 species, and of GCase proteins.

• Journal of Ginseng Research
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• v.9 no.1
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• pp.54-63
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• 1985

The free phenolic fraction from Korean white ginseng (Panax ginseng C.A. Meyer) was studied by GLC/MS as trimethylsilyl and methyl derivative. Five phenolic compounds such as 2,6-ditert butyl p-crestal, phloroglucinol, protocatechuic acid, isoferulic acid, quinic acid were identified newly. And additionally 13 organic acids and hydrocarbons were also identified in the fraction.

• Korean Journal of Environmental Agriculture
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• v.4 no.2
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• pp.114-117
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• 1985

A new method for micro-quantification of abscisic acid from plant tissue is described. GLC-ECD method of abscisic acid pertafluorobenzyl ester synthesized by reaction of plant extract with pentafluorobenzyl bromide showed higher sensitivity than that of abscisic acid methyl ester, and the detection limit of abscisic acid by the described method was as low as 5 pg.

• Journal of Applied Biological Chemistry
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• v.48 no.4
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• pp.218-221
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• 2005

Nucleotide sequence of Brevibacterium flavum ptsG gene capable of complementing Escherichia coli ZSC113 mutations defective to glucose permease activity of phosphotransferase system was completely determined, and the gene product was compared with other glucose-specific enzyme II ($EII^{Glc}$). A ptsG gene of B. flavum consisted of open reading frame of 2,025 nucleotides putatively encoding polypeptide of 675 amino acid residues and TAA stop codon. Deduced amino acid sequence of B. flavum ($EII^{Glc}$) had high homology with ($EIIs^{Glc}$) of Corynebacterium glutamicum, C. efficiens, and B. lactofermentum. Arrangement of structural domains, IIBCA, of B. flanum ($EII^{Glc}$) protein was identical to that of EIIs belonging to glucose-phosphotransferase system.

• Journal of Life Science
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• v.2 no.4
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• pp.232-239
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• 1992

UDP_GlcNAc is metabolized to form vegetative cell wall, cortical peptidoglycans, and outermost layer consisting of galactosamine-6-phosphate ploysaccharide in life cycle of Bacillus megaterium. To obtain a better understanding of the UDP-GlcNAc regulation, we examined the activity of the common first enzyme for the synthesis of nucleotide precursors of peptidoglycans, enoylpyruvate transferase by newly developed method. Both the specific and the total activity decreased after the end of exponential growth followed by and increase from t5 but decreased again parallel to the appearance of the activity of UDP_GlcNAc-4-epimerase. Antibody specificity to anti-transferase IgG and the elution profile on DEAE-Sepharose revealed that B. megaterium has at least two enoylpyruvate transferase isozymes, and UDP_GlcNAc was metabolized to vegetative cell wall and cortical peptidoglycan by each isozme in exponential growth and in sporulation, respectively in life cycle.

• Bulletin of the Korean Chemical Society
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• v.29 no.7
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• pp.1315-1319
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• 2008

A trisaccharide, 6d-Altro-Hepp$\alpha$ (1$\rightarrow$3) GlcNAc$\beta$ (1$\rightarrow$3) Gal$\alpha$ (1$\rightarrow$$OCH_2CH_2N_3$, as an O-antigenic repeating unit of Campylobacter jejuni serotypes O:23 and O:36, was synthesized. Coupling of the 6d-altro-Hepp$\alpha$ (1$\rightarrow$3) GlcNAc$\beta$ (1$\rightarrow$SEt donor with Gal$\alpha$ (1${\rightarrow}OCH_2CH_2Cl$ acceptor in the presence of NIS-TfOH promoter afforded the trisaccharide having the $\beta$ (1$\rightarrow$3) Gal linkage. $\beta$ -Stereospecificity and the desired regioselectivity for the 3-OH Gal are obtained. Subsequent hydrogenation, acetylation, azide displacement, hydrazinolysis, Nacetylation, and finally deacetylation furnished the title trisaccharide hapten for further glycoconjugation.