• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.3
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• pp.192-198
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• 2003

BiP, immunoglobulin binding protein, is an ER homologue of Hsp70. However, unlit other Hsp70 proteins, regulatory protein(s) for BiP has not been identified. Here, we demo strafed the presence of potential regulatory proteins for BiP using a pull -down assay. Since BiP can bind any unfolded protein, only the ATPase domain of BiP was used for the pull -down assay in order to minimize nonspecific binding. The ATPase domain was cloned to produce recombinant protein, which was then conjugated to CNBr-activated agarose. The structural conformation and ATP hydrolysis activity of the recombinant ATPase domain were similar to those of the native protein, light proteins from metabolically labeled mouse plasmacytoma cells specifically bound to the recombinant ATPase protein. The binding of these proteins was inhibited by excess amounts of free ATPase protein, and was dependent on the presence of ATP. These proteins were eluted by ADP. Of these proteins, Grp170 and BiP where identified. while the other were not identified as known ER proteins, from Western blot analyses. The presence of the ATPase-binding proteins for BiP was first demonstrated in this study, and our data suggest similar regulatory machinery for BiP may exist in the ER, as found in prokaryotes and other cellular compartments.

• The Plant Pathology Journal
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• v.33 no.2
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• pp.148-162
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• 2017

As a step towards discovering novel pathogenesis-related proteins, we performed a genome scale computational identification and characterization of secreted and transmembrane (TM) proteins, which are mainly responsible for bacteria-host interactions and interactions with other bacteria, in the genomes of six representative Burkholderia species. The species comprised plant pathogens (B. glumae BGR1, B. gladioli BSR3), human pathogens (B. pseudomallei K96243, B. cepacia LO6), and plant-growth promoting endophytes (Burkholderia sp. KJ006, B. phytofirmans PsJN). The proportions of putative classically secreted proteins (CSPs) and TM proteins among the species were relatively high, up to approximately 20%. Lower proportions of putative type 3 non-classically secreted proteins (T3NCSPs) (~10%) and unclassified non-classically secreted proteins (NCSPs) (~5%) were observed. The numbers of TM proteins among the three clusters (plant pathogens, human pathogens, and endophytes) were different, while the distribution of these proteins according to the number of TM domains was conserved in which TM proteins possessing 1, 2, 4, or 12 TM domains were the dominant groups in all species. In addition, we observed conservation in the protein size distribution of the secreted protein groups among the species. There were species-specific differences in the functional characteristics of these proteins in the various groups of CSPs, T3NCSPs, and unclassified NCSPs. Furthermore, we assigned the complete sets of the conserved and unique NCSP candidates of the collected Burkholderia species using sequence similarity searching. This study could provide new insights into the relationship among plant-pathogenic, humanpathogenic, and endophytic bacteria.

• Journal of Microbiology and Biotechnology
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• v.24 no.6
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• pp.854-861
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• 2014

The diagnosis of Brucella abortus is mainly based on serological methods using antibody against LPS, which has diagnostic problems. Therefore, to solve this problem, we evaluated two proteins of B. abortus, Cu/Zn superoxide dismutase (SodC) and outer membrane proteins 2b porin (Omp2b). The genes were cloned and expressed in a pMAL system, and the recombinant proteins, rOmp2b and rSodC, were purified as fusion forms with maltose-binding protein. The identity of the proteins was confirmed by SDS-PAGE and Western blot analysis with sera of mice infected with B. abortus. Production of cytokines and nitric oxide (NO) was investigated in RAW 264.7 cells and mouse splenocytes after stimulation with the proteins. Moreover, cellular and humoral immune responses were investigated in BALB/c mice after immunization with the proteins. TNF-${\alpha}$, IL-6, and NO were significantly inducible in RAW 264.7 cells. Splenocytes of naive mice produced IFN-${\gamma}$ and IL-4 significantly by stimulation. Moreover, number of IgG, IFN-${\gamma}$, and IL-4 producing cells were increased in immunized mice with the two proteins. Production of IgG and IgM with rOmp2b was higher than those with rSodC in immunized mice. These results suggest that the two recombinant proteins of B. abortus may be potential LPS-free proteins for diagnosis.

• Journal of Microbiology
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• v.39 no.3
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• pp.162-167
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• 2001

Methylovorus sp. strain SS1 DSM 11726 was found to grow continuously when it was transferred from 30$\^{C}$ to 40$\^{C}$ and 43$\^{C}$. A shift in growth temperature from 30$\^{C}$ to 45$\^{C}$, 47$\^{C}$ and 50$\^{C}$ reduced the viability of the cell population by more than 10$^2$, 10$^3$and 10$\^$5/ folds, respectively, after 1h cultivation. Cells transferred to 47$\^{C}$ and 50$\^{C}$ after preincubation for 15 min at 43$\^{C}$, however, exhibited 10-fold increase in viability. It was found that incubation for 15 min at 40$\^{C}$ of Methylovorus sp. strain SSl grown at 30$\^{C}$ was sufficient to accelerate the synthesis of a specific subset of proteins. The major heat shock proteins had apparent molecular masses of 90, 70, 66, 60, and 58 kDA. The 60 and 58 kDa proteins were found to cross-react with the antiserum raised against GroEL protein. The heat shock response persisted for over 1h. The shock proteins were stable for 90 min in the cell. Exposure of the cells to methanol induced proteins identical to the heat shock proteins. Addition of ethanol induced a unique protein with a molecular mass of about 40 kDa in addition to the heat-induced proteins. The proteins induced in paraquat-treated cells were different from the heat shock proteins, except the 70 and 60 kDa proteins.

• BMB Reports
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• v.48 no.9
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• pp.531-536
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• 2015

Gecko proteins have long been used as anti-tumor agents in oriental medicine, without any scientific background. Although anti-tumor effects of Gecko proteins on several cancers were recently reported, their effect on bladder cancer has not been investigated. Thus, we explored the anti-tumor effect of Gecko proteins and its cellular mechanisms in human bladder cancer 5637 cells. Gecko proteins significantly reduced the viability of 5637 cells without any cytotoxic effect on normal cells. These proteins increased the Annexin-V staining and the amount of condensed chromatin, demonstrating that the Gecko proteinsinduced cell death was caused by apoptosis. Gecko proteins suppressed Akt activation, and the overexpression of constitutively active form of myristoylated Akt prevented Gecko proteins-induced death of 5637 cells. Furthermore, Gecko proteins activated caspase 9 and caspase 3/7. Taken together, our data demonstrated that Gecko proteins suppressed the Akt pathway and activated the intrinsic caspase pathway, leading to the apoptosis of bladder cancer cells. [BMB Reports 2015; 48(9): 531-536]

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.126-126
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• 2017

Mitochondria are important in wheat, as in all crops, as the main source of ATP for cell maintenance and growth including vitamin synthesis, amino acid metabolism and photorespiration. To investigate the mitochondrial proteome of the roots of wheat seedlings, a systematic and targeted analysis were carried out on the mitochondrial proteome from 15 day-old wheat seedling root material. Mitochondria were isolated by Percoll gradient centrifugation; and extracted proteins were separated and analyzed by Tricine SDS-PAGE along with LTQ-FTICR mass spectrometry. From the isolated the sample, 184 proteins were identified which is composed of 140 proteins as mitochondria and 44 proteins as other subcellular proteins that are predicted by the freeware subcellular predictor. The identified proteins in mitochondria were functionally classified into 12 classes using the ProtFun 2.2 server based on biological processes. Proteins were shown to be involved in amino acid biosynthesis (17.1%), biosynthesis of cofactors (6.4%), cell envelope (11.4%), central intermediary metabolism (10%), energy metabolism (20%), fatty acid metabolism (0.7%), purines and pyrimidines (5.7%), regulatory functions (0.7%), replication and transcription (1.4%), translation (22.1%), transport and binding (1.4%), and unknown (2.8%). These results indicate that many of the protein components present and functions of identifying proteins are common to other profiles of mitochondrial proteins performed to date. This dataset provides the first extensive picture, to our knowledge, of mitochondrial proteins from wheat roots. Future research is required on quantitative analysis of the wheat mitochondrial proteomes at the spatial and developmental level.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.120-120
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• 2017

Since the composition of proteins from the Korean cultivars of Proso millet is unknown; thereby, the present study was conducted to obtain a reference map of millet seed proteins and identify the functional characteristics of the identified proteins. Proteins extracted from the millet seeds of various cultivars, were investigated using proteomic techniques as 2D electrophoresis coupled with mass fingerprinting. The 1152 (differentially expressed) proteins were detected on 2-D gel. Among them, 26 reproducible protein spots were analyzed by MALDI-TOF-TOF mass spectrometry. Out of 26 proteins, 2 proteins were up-regulated towards all cultivars of millet, while 7 proteins were up-regulated and 13 proteins were down-regulated against only one cultivar. However, abundance in most identified protein species, associated with metabolism, transcription and transcription was significantly enhanced, while that of another protein species involved in polysaccharide metabolism, stress response and pathogenesis were severely reduced. Taken together, the results observed from the study suggest that the differential expression of proteins from the four cultivars of millet may be cultivar-specific. Taken together, a proteomic investigation of millet seeds from different cultivars, we sought to better understand the genetic variation of millet cultivars representing the future millet research, and the functional categorization of individual proteins on the basis of their molecular function.

• Journal of Nutrition and Health
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• v.2 no.4
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• pp.127-134
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• 1969

Total and esterified cholesterol content was determined in the rat administered animal and vegetable proteins for 16 weeks. The cholesterol biosynthetic activity of the liver was also measured in these rats by the $acetate-C^{14}$ incorporation rate. The results obtained were as follows. (1) Serum total cholesterol content was increased by the administration of animal proteins and decreased by that of vegetable proteins. (2) Liver cholesterol content was increased by animal proteins and decreased by at of vegetable proteins. (3) Cholesterol biosynthetic activity of the liver was increased by the animal proteins and decreased by the vegetable proteins.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.2
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• pp.496-499
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• 2004

We have previously observed that Bambusae Caul is in Liquamen (BCL) stimulates the adipose conversion of 3T3-L1 cells and molecular chaperones were involved in the process of the assembly and replacement of laminin subunits in Bovine aortic endothelial cells(BAEC). Endothelial cells are exposed to continuous shear stress due to the blood flow. Heat shock protens(hsp) are a well-known stress response protein, namely, stress proteins. To investigate effects of BCL on the stress proteins induced by heating in endothelial cells, we have analyzed synthetic amounts of stress proteins in sodium dodecyl sulfate gel electrophoresis under reducing conditions. Under the condition of heating stress, BCL inhibited the synthesis of stress proteins in endothelial cells. These results suggest that BCL may have an important role for expression of stress proteins induced by heating in endothelial cells.

• Microbiology and Biotechnology Letters
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• v.25 no.2
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• pp.218-223
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• 1997

For the utilization of animal blood produced in slaughter for the cultivation of lactic acid bacteria, the nitrogen sources in a complex(MRS) medium were replaced by blood plasma proteins. Focusing the purpose on the industrial production of a probiotics, the hydrolytic activities of three industrially applicable proteases were compared for the effective digestion of the proteins, and Alcalase(the product of Novo Nordisk) was selected with comparatively high activity. The growth of Streptococcus thermophilus KCCM12020 was best among the four strains of lactic acid bacteria tested. With Alcalase-digested proteins in the medium, the growth rates and the final cell concentrations were higher than those with non-digested proteins. The cell mass produced in the medium containing blood proteins as nitrogen sources, $2.5{\times}10^9$ CFU/ml, was significantly high and about 70% of that in MRS medium, showing a great possibility for the utilization of animal blood proteins as economic nitrogen sources in the production of cell mass of lactic acid bacteria.