• Korean Journal of Environmental Biology
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• v.20 no.1
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• pp.20-24
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• 2002

We have from water samples of Kwalim, Dochang, and Mulwang reservoirs in Kyonggi-Do, where cyanobacteria blooming occurred. Isolated microbes which have lytic activity for cyanobacteria. Water samples were smeared on the Anabaena cylindrica lawn and incubated in light chamber at $28^\circ{C}$, under 3000 lux for 13 days. A fungus having cyanobacterial lytic activity was isolated from the samples of Dochang reservoir. The isolate was identified as Cryptococcus laurentii by Vitek system. From the culture of the isolate, four major extracellular protein bands (29, 35.2, 40.9, 51.1 kDa) have been detected and the 29 kDa protein band was more thickly appeared in the culture with cyanobacteria.

• Applied Biological Chemistry
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• v.36 no.1
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• pp.11-16
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• 1993

A bacteria strain producing extracellular protein was isolated and identified from soil samples, and the optimum conditions of producing protein were investigated. Eleven strains of bacteria were isolated from soil samples. Among which WY-60 strain showed a very strong capability of producing protein and identified as a Bacillus sp. The optimum composition of nutrient medium for the production of the protein by WY-60 was fructose 4.0%, polypeptone 1.0%, $NH_4NO_3$ 0.1%, $K_2HPO_4$ 0.1%, $MgSO_4{\cdot}7H_2O$ 0.005%, $CaCO_3$ 1.0% and the optimum pH and temperature were 8.0 and $30^{\circ}C$, respectively. The penicillin G and lincomycin added to the above medium were effective for the protein production of the WY-60, but other antibiotics were non-effective. The maximum production of protein was obtained after 5 days culture.

• Journal of Life Science
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• v.20 no.8
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• pp.1166-1172
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• 2010

Kinesin-I exists as a tetramer of two heavy chains (KHCs, also called KIF5s), which contain the amino (N)-terminal motor domain and carboxyl (C)-terminal domain, as well as two light chains (KLCs), which bind to the KIF5s (KIF5A, KIF5B and KIF5C) stalk region. To identify the interaction proteins for KIF5A, yeast two-hybrid screening was performed and a specific interaction with the ${\beta}$ subunit of heterotrimeric G proteins ($G{\beta}$) was found. $G{\beta}$ bound to the amino acid residues between 808 and 935 of KIF5A and to other KIF5 members in the yeast two-hybrid assay. The WD40 repeat motif of $G{\beta}$ was essential for interaction with KIF5A. In addition, these proteins showed specific interactions in the glutathione S-transferase (GST) pull-down assay. An antibody to KIF5s specifically co-immunoprecipitated KIF5s associated with heterotrimeric G proteins from mouse brain extracts. These results suggest that kinesin-I motor protein transports heteroterimeric G protein attachment vesicles along microtubules in the cell.

• Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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• 1993.12a
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• pp.10-11
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• 1993

기저상태의 산소분자는 비교적 안정하지만 생체ㅇ내외에서 물리적, 화학적으로 활성화 되어 $O_{2}$, $^{1}O_{2}$, OH, $H_{2}O_{2}$ 등의 활성산소종을 생성하며, 생체의 지질, 단백질, 핵산 당등의 분자에 산화적 상해를 초래하여 노화, 암, 순환기, 호흡기 게통의 질환과 식품의 품질열화에 관여하는 것으로 알려져 있다. 따라서 본인은 식품의 맛, 향기, 색 등을 부여하는 고유의 기능 외에도 방부제, 한방약으로 널리 이용되고 오고 있는 51종의 향신료를 대상으로 활성산소포촉활성을 조사하고 나아가 활성물질을 분리, 정제 및 동정함으로 향신료의 새로운 기능을 밝히고 신약 개발의 기초적 자료를 제시하고저 한다. Fenton 반응을 이용하여 2-deozyribose 산화법과 sodium benzoic acid 수산화법으로 51종의 향신료의 OH 포촉활성을 검색한 결과, Cruciferae과의 nustard 류, Labiatae과의 thyme, saga, savory, oregano, Myrtaceae과의 clove, allspice 가 1ug/ml 농도에서 50%이상의 포촉활성을 나타내었으며 그중 mustard 류는 같은 농도에서 거의 90%이상의 활성을 나타내었다. 활성물질의 분리 및 정제는 Amberlite XAD-2 갈럼과 preparative-HPLC를 이용 하였으며, EI, FAB-MS, IR, $^{1}H$, $^{13}C-NMR$, Cosy-NMR로 그 화학적 구조를 동정하였다. Brown mustard에서 동정된 4-hydroxy-3,5-dimethoxy cinnamic acid Methyl ester는 0.42$\mu$ mol 농도에서 90% 이상의 OH 포촉활성을 나타내어 이를 diazomethane 반응으로 조제하였으며 white mistard에서는 4-hydroxy-3,5-dimethoxy cinnamoyl choline을 동정하였다.

• Journal of Plant Biotechnology
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• v.50
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• pp.1-10
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• 2023

Sweet potato (Ipomoea batatas L. Lam) is an economically important root crop and a valuable source of nutrients, processed foods, animal feeds, and pigment materials. However, during post-harvest storage, storage roots of sweet potatoes are susceptible to decay caused by various microorganisms and diseases. Post-harvest curing is the most effective means of healing wounds and preventing spoilage by microorganisms during storage. In this study, we aimed to identify proteins involved in the molecular mechanisms related to curing and study proteomic changes during the post-curing storage period. For this purpose, changes in protein spots were analyzed through 2D-electrophoresis after treatment at 33℃ (curing) and 15℃ (control) for three days, followed by a storage period of eight weeks. As a result, we observed 31 differentially expressed protein spots between curing and control groups, among which 15 were identified. Among the identified proteins, the expression level of 'alpha-amylase (spot 1)' increased only after the curing treatment, whereas the expression levels of 'probable aldo-keto reductase 2-like (spot 3)' and 'hypothetical protein CHGG_01724 (spot 4)' increased in both the curing and control groups. However, the expression level of 'sporamin A (spot 10)' decreased in both the curing and control treatments. In the control treatment, the expression level of 'enolase (spot 14)' increased, but the expression levels of 'chain A of actinidin-E-64 complex+ (spot 19)', 'ascorbate peroxidase (spot 22)', and several 'sporamin proteins (spot 20, 21, 23, 24, 27, 29, 30, and 31)' decreased. These results are expected to help identify proteins related to the curing process in sweet potato storage roots, understand the mechanisms related to disease resistance during post-harvest storage, and derive candidate genes to develop new varieties with improved low-temperature storage capabilities in the future.

• Korean Journal of Microbiology
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• v.41 no.4
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• pp.262-268
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• 2005

Intoxication of murine macrophages (RAW 264.7) with the anthrax lethal toxin (LeTx 100 ng/ml) results in profound alterations in the host cell gene expression. The role of LeTx in mediating these effects is unknown, largely due to the difficulty in identifying and assigning function to individual proteins. In this study, we have used two-dimensional polyacrylamide gel electrophoresis to analyze the protein profile of murine macrophages treated with the LeTx, and have coupled this to protein identification using MALDI-TOF mass spectrometry. Interpretation of the peptide mass fingerprint data has relied primarily on the ProFound database. Among the differentially expressed spots, cleaved mitogen-activated protein kinase kinase (Mek1) and glucose-6-phosphate dehydrogenase were increased in the LeTx treated macrophages. Mek1 acts as a negative element in the signal transduction pathway, and G6PD plays the role for the protection of the cells from the hyper-production of active oxygen. Our results suggest that this proteomic approach is a useful tool to study protein expression in intoxicated macrophages and will contribute to the identification of a putative substrate for LeTx.

• Applied Biological Chemistry
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• v.35 no.2
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• pp.132-138
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• 1992

For effective determination of methylthiohydantoin amino acids(MTHs) by gas liquid chromatography in the protein sequencing, derivatization with N-methyl-N-(tert.-butyl-dimethylsilyl)trifluoroacetamide(MTBSTFA), a new silylating reagents, was attempted instead of trimethylsilyl(TMSi) derivatives by N,O-bis(trimethylsilyl)trifluoroacetamide(BSTFA) used up to the present and N(O)-butyldimethylsilyl MTHs derivatized by MTBSTFA were analysed on HP-1 capillary column. Twenty one protein amino acids except cystine were indentified. Especially arginine that did not detected with TMSi derivative on packed column until now was resolved by derivatization with MTBSTFA. N(O)-butyldimethylsilyl MTHs showed multiple peaks by MTBSTFA were proline, isoleucine, glycine and tyrosine and hydroxyproline especially showed several extraneous peaks more than two. Calibration curves of N(O)-butyldimethysilyl MTHs of amino acids in the range of $2.5\;nmol{\sim}7.5\;nmole$ showed good linearity. however, those of lysine, histidine and arginine showed linearity in the range of $5.0\;nmole{\sim}15.0\;nmole$. Correlation coefficients and regression coefficients of all calibration curves were highly significant(p<0.001).

• Microbiology and Biotechnology Letters
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• v.35 no.2
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• pp.112-117
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• 2007

Protein kinases play very important role for maintaining viability in prokaryote and eukaryote. The metabolism of prokaryotic cell is generally regulated by bacterial two-component regulatory systems that are composed of histidine and asparitic acid kinases, however, some eukaryotic signal transduction system such as, serine and threonine kinases, have been also found to be involved in the regulation of morphogenesis and physiological differentiation in Streptomyces. Streptomyces griseus, a streptomycin producer, was expected to have varlous types of eukaryotic-type serine/threonine protein kinases, controlling morphogenesis. Thus, many steps of chromatographies were applied to isolate serine and threonine kinases from S. griseus IFO13350. The immunoaffinity steps using anti-phosphoserine, anti-phosphothreonine, and anti-phosphotyrosine agarose column chramatographies were successfully introduced to identify eukaryotic protein kinases from S. griseus IFO13350. Eight proteins with the expected molecular weight of 14, 29, 31, 35, 40, 52, 56, and 60 kDa, were identified on SDS-PAGE, and the their kination activity was confirmed by nonradioactive protein kination assay using FITC-labeled peptide as the substrate.

• Applied Biological Chemistry
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• v.27 no.2
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• pp.129-134
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• 1984

Change of protein component in soybean sprout grown at four temperatures was investigated by polyacrylamide gel electrophoresis. Main bands were identified using purified seed globulins. Electrophoretogram showed 5 main bands (a. b, c, d, and p) and 10 minor bands in seed and maximum number (19) of bands (8 main band including 0 and 11 minor) at 4th day after germination in cotyledon. All bands appeared in axis protein but resolution was poor. In cotyledon, a component (most rapidly) and b+c+d component decreased while o+p component and other minor components were increased at 6th day and decreased thereafter. In axis all components increased rapidly, especially in minor components and b+c+d component. High growing temperature accelerated decrease in cotyledon and increase in axis of protein, especially for 11S. The a component was identified as 7S, b+c+d as 11S and o+p as 2S globulin.

• Journal of Life Science
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• v.14 no.1
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• pp.26-31
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• 2004

Redox factor-1 (Ref-1), known as a redox regulator, controls the DNA binding of AP-1 and is activated in HT29 colon cancer cells by hypoxia in vitro. REF-1 also increases tile DNA binding affinity of Hypoxia-inducible Factor-lalpha$ (HIF-lalpha$), HIF-like Factor (HLF) and early growth response-1 (Egr-1) which induce expression of the genes involved in angiogenesis, so that we speculate that REF-1 may play a role in hypoxia-induced angiogenesis. In this research we tried to detect novel proteins interacting with REF-1 using Yeast two-hybrid system using full-length REF-1 cDNA as bait. As result of such screening we detected 3 positive clones. DNA sequencing and GeneBank search revealed that one of the clones contained the same sequences as M.musculus cDNA for tioredoxin.