• Proceedings of the Korean Journal of Food and Nutrition Conference
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• 2001.12a
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• pp.121-121
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• 2001

최소 배지에 여러 아미노산과 대사 산물을 첨가하여 배양시킨 Serratia marcescens ATCC 25419 세포추출물에서여 biodegradative threonine dehydratase (BDTD), biosynthetic threonine dehydratase (BSTD)와 acetolactate syntase (ALS)의 비활성도를 조사하였다. S. marcescens BDTD와 ALS는 낮은 농도 (0.5-2 mM)의 cAMP에 의해 촉진적 조절을 받으며, 비교적 낮은 농도의 isoleucine (1-4 mM)에 의해서는 S. marcescens BSTD의 생합성이 증가되고 높은 농도의 isoleucine (10-30 mM)에서는 감소되고 비교적 낮은 농도의 valine (2-4 mM)에 의해서 S. marcescens ALS의 생합성이 증가되는 것으로 보아 S. marcescens ATCC 25419에서 branched chain 아미노산 생합성 과정의 조절 양상은 Escherichia coli K-12와는 달리, isoleucine의 생합성 과정은 BSTD에 의해 조절되고, valine의 생합성 과정은 ALS에 의해 조절되는 것으로 사료된다.

• Proceedings of the Botanical Society of Korea Conference
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• 2002.04a
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• pp.73-82
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• 2002

The committing step in Met and S-adenosyl-L-methionine (SAM) synthesis is catalyzed by cystathionine ${\gamma}$ -synthase (CGS). Transgenic Arabidopsis thaliana overexpressing CGS under control of 35S promoter show increased soluble Met and its metabolite S-methylmethionine, but only at specific stages of development. CGS-overexpressing seedlings are resistant to ethionine. Similar results were obtained with transgenic potato plants overexpressing Arabidopsis CGS. Several of the transgenic lines show silencing of CGS resulting in deformed p]ants with a reduced capacity for reproductive growth similar as transgenic plants by antisense RNA (CGS[-]). Exogenous feeding of Met to the CGS[-] and CGS[+] silenced plants partially restores their growth. Similar morphological deformities are observed in plants cosuppressed for SAM synthetase, even though such plants accumulate 250 fold more soluble Met than wild type and they overexpress CGS. The results suggest that the abnormalities associated with CGS and SAM synthetase silencing are due in part to a reduced ability to produce SAM, and that SAM may be a regulator of CGS expression.

• Journal of Integrative Natural Science
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• v.8 no.1
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• pp.40-47
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• 2015

Glycogen synthase kinase 3 (GSK-3) is a serine/threonine protein kinase that has recently emerged as a promising target in drug discovery. It is involved in multiple cellular processes and associated with the pathogenesis of several diseases. A three-dimensional quantitative structure-activity relationship (3D-QSAR) analysis was performed on a series of GSK-3 inhibitors to understand the structural basis for inhibitory activity. Comparative molecular field analysis (CoMFA) method was used to derive 3D-QSAR models. A reliable CoMFA model was developed using ligand-based alignment scheme. The model produced statistically acceptable results with a cross-validated correlation coefficient ($q^2$) of 0.594 and a non-cross-validated correlation coefficient ($r^2$) of 0.943. Robustness of the model was checked by bootstrapping and progressive scrambling analysis. This study could assist in the design of novel compounds with enhanced GSK-3 inhibitory activity.

• KSBB Journal
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• v.7 no.3
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• pp.155-160
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• 1992

To improve the nutritional quality of plant proteins, a synthetic gene, called HEAAE (high essential amino acid encoding)-DNA, was introduced and expressed in tobacco plants. The synthetic gene, which is 292 basepair-long, codes for a protein composed of about 80% essential amino acids. To improve its expression level in plants, Cauliflower Mosaic Virus (CaMV) 355 and CaMV duplicate 35S promoters which are known as strong promoters were used with Nopaline Synthase promoter as a control. Transformed and regenerated tobacco plants were subject to analysis for introduction and expression of this gene. Integration of the gene into the plant genome and its expression into mRNAs and its proteins have been demonstrated using Southern, northern blot analysis and amino acid analysis. The differences of expression levels among CaMV duplicate 35S, CaMV 35S and Nopaline Synthase promoters are significant in term of mRNAs, but not in terms of proteins.

• BMB Reports
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• v.49 no.1
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• pp.51-56
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• 2016

Glycogen synthase kinase-3β (GSK-3β) is a serine/threonine protein kinase that is known to mediate cancer cell death. Here, we show that B-cell lymphoma 2 (Bcl-2), an anti-apoptotic protein, is regulated by GSK-3β and that GSK-3β-mediated regulation of Bcl-2 is crucial for mitochondrial-dependent cell death in paclitaxel-stimulated cells. We demonstrate that MCF7 GSK-3β siRNA cells are more sensitive to cell death than MCF7 GFP control cells and that in the absence of GSK-3β, Bcl-2 levels are reduced, a result enhanced by paclitaxel. Paclitaxel-induced JNK (c-Jun N-terminal kinase) activation is critical for Bcl-2 modulation. In the absence of GSK-3β, Bcl-2 was unstable in an ubiquitination-dependent manner in both basal- and paclitaxel-treated cells. Furthermore, we demonstrate that GSK-3β-mediated regulation of Bcl-2 influences cytochrome C release and mitochondrial membrane potential. Taken together, our data suggest that GSK-3β-dependent regulation of Bcl-2 is crucial for mitochondria-dependent cell death in paclitaxel-mediated breast cancer therapy. [BMB Reports 2016; 49(1): 51-56]

• The Plant Pathology Journal
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• v.19 no.5
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• pp.239-247
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• 2003

In Korea, a local soybean (Glycine max) genotype 56l. was found to be strongly resistant to a virulent bacterial strain of a Pseudomonas sp. SN239. Specific genes involved in the resistance of the soybean genotype 561 were identified and the pattern of gene expression against the Pseudomonas infection was analyzed using differential-display reverse transcription PCR (DDRT-PCR). More than 126 cDNA fragments representing mRNAs were induced within 48 hours of bacteria inoculation. Among them, 28 cDNA fragments were cloned and sequenced. Twelve differentially displayed clones with open reading frames had unknown functions. Sixteen selected cDNA clones were homologous to known genes of other organisms. Some of the identified cDNAs were pathogenesis-related (PR) genes and PR-like genes. These cDNAs included a putative calmodulin-binding protein; an endo-l,3-1,4-$\bate$-D-glucanase; a $\bate$-1,3-endoglucanase; a $\bate$-1,3-exoglucanase; a phytochelatin synthetase-like gene; a thiol protease; a cycloartenol synthase; and a putative receptor-like serine/threonine protein kinase. Among them, four genes were found to be putative PR genes induced significantly by the Pseudomonas infection. These included a calmodulin-binding protein gene, a $\bate$-1,3-endoglucanase gene, a receptor-like serine/threonine protein kinase gene, and pS321 (unknown function). These results suggest that the differentially expressed genes may mediate the strong resistance of soybean 561 to the strain SN239 of Pseudomonas sp.

• Molecules and Cells
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• v.26 no.1
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• pp.100-105
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• 2008

Glycogen synthase kinase $3{\beta}$ (GSK $3{\beta}$) is a serine/threonine kinase that phosphorylates substrates such as ${\beta}$-catenin and is involved in a variety of biological processes, including embryonic development, metabolism, tumorigenesis, and cell death. Here, we present evidence that human GSK $3{\beta}$ is associated with Fe65, which has the characteristics of an adaptor protein, possessing a WW domain, and two phosphotyrosine interaction domains, PID1 and PID2. The GSK $3{\beta}$ catalytic domain also contains a putative WW domain binding motif ($^{371}PPLA^{374}$), and we observed, using a pull down approach and co-immunoprecipitation, that it interacts physically with Fe65 via this motif. In addition, we detected co-localization of GSK $3{\beta}$ and Fe65 by confocal microscopy, and this co-localization was disrupted by mutation of the putative WW domain binding motif of GSK $3{\beta}$. Finally, in transient transfection assays interaction of GSK $3{\beta}$ (wt) with Fe65 induced substantial cell apoptosis, whereas interaction with the GSK $3{\beta}$ AALA mutant ($^{371}AALA^{374}$) did not, and we noted that phosphorylation of the Tyr 216 residue of the GSK $3{\beta}$ AALA mutant was significantly reduced compared to that of GSK $3{\beta}$ wild type. Thus, our observations indicate that GSK $3{\beta}$ binds to Fe65 through its $^{371}PPLA^{374}$ motif and that this interaction regulates apoptosis and phosphorylation of Tyr 216 of GSK $3{\beta}$.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.10 no.7
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• pp.1766-1772
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• 2009

In this study, we are interested in comparing the protein profiles of acid-shocked and control cells of S. mutans isolated from Korean children with caries. The results of 2D gel electrophoresis showed that twelve proteins are up-regulated when the cells were grown under 20 mM lactic acid stress in the exponential phase. Up-proteins under acid stress were estimated a major key of the survival and proliferation of S. mutans in low pH environments. These proteins are estimated generally associated with three biochemical pathways: glycolysis, alternative acid production and branched-chain amino acid biosynthesis.

• The Korean Journal of Pesticide Science
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• v.7 no.4
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• pp.239-247
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• 2003

This experiment was carried out to study the resistant mechanism of sulfonylurea(SU) herbicides to Monochoria korsakowii occurring in the rice fields of Korea. The activity of acetolactate synthase(ALS), absorption and translocation of $[^{14C}]$bensulfuron-methyl, and DNA sequence of ALS genes were studied. The apparent SU resiatance to Monochoria korsakowii was confirmed in greenhouse testes. Fresh weight accumulation$(GR_{50})$ in the resistant biotype was about 5- to 64-fold higher in the presence of six SU herbicides compared to the susceptible biotype. The ALS activity isolated from the resistant biotype to herbicides tested was less sensitive than that of susceptible biotype. The concentration of herbicide required for 50% inhibition of ALS activity$(I_{50})$ was 14- to 76-fold higher as compared to the susceptible biotype. No differences were observed in the rates of $[^{14C}]$bensulfuron uptake and translocation. However, the DNA sequence from the resistant biotype differed from that of the susceptible biotype by single nucleotide substitution at three amino acid each in the middle region excluding the ends of ALS genes. We found three point mutations causing substitution of serine for threonine at amino acid 168, arginine for histidine at amino acid 189, and a aspartic acid for phenylalanine at amino acid 247, respectively, in the resistant biotype.

• Biomolecules & Therapeutics
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• v.31 no.1
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• pp.127-138
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• 2023

Glycogen synthase kinase-3β (GSK-3β) is an important serine/threonine kinase that implicates in multiple cellular processes and links with the neurodegenerative diseases including Alzheimer's disease (AD). In this study, structure-based virtual screening was performed to search database for compounds targeting GSK-3β from Enamine's screening collection. Of the top-ranked compounds, 7 primary hits underwent a luminescent kinase assay and a cell assay using human neuroblastoma SH-SY5Y cells expressing Tau repeat domain (Tau_RD) with pro-aggregant mutation ΔK280. In the kinase assay for these 7 compounds, residual GSK-3β activities ranged from 36.1% to 90.0% were detected at the IC₅₀ of SB-216763. In the cell assay, only compounds VB-030 and VB-037 reduced Tau aggregation in SH-SY5Y cells expressing ΔK280 Tau_RD-DsRed folding reporter. In SH-SY5Y cells expressing ΔK280 Tau_RD, neither VB-030 nor VB-037 increased expression of GSK-3α Ser21 or GSK-3β Ser9. Among extracellular signal-regulated kinase (ERK), AKT serine/threonine kinase 1 (AKT), mitogen-activated protein kinase 14 (P38) and mitogenactivated protein kinase 8 (JNK) which modulate Tau phosphorylation, VB-037 attenuated active phosphorylation of P38 Thr180/ Tyr182, whereas VB-030 had no effect on the phosphorylation status of ERK, AKT, P38 or JNK. However, both VB-030 and VB-037 reduced endogenous Tau phosphorylation at Ser202, Thr231, Ser396 and Ser404 in neuronally differentiated SH-SY5Y expressing ΔK280 Tau_RD. In addition, VB-030 and VB-037 further improved neuronal survival and/or neurite length and branch in mouse hippocampal primary culture under Tau cytotoxicity. Overall, through inhibiting GSK-3β kinase activity and/or p-P38 (Thr180/Tyr182), both compounds may serve as promising candidates to reduce Tau aggregation/cytotoxicity for AD treatment.