• 한국유기농업학회지
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• 제22권1호
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• pp.167-182
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• 2014

Korean Panax ginseng C. A. Meyer (P. ginseng) is a well-known and one of the most important tonic herbs used in traditional Korean medicine. The pharmacological effects of P. ginseng have been reported by many researchers. Nevertheless, little is known between the mechanism of action and the active compounds. In this study, we performed a comprehensive proteomic analysis and protein categorization in order to understand the physiological characteristics of the major components in the adventitious roots of P. ginseng. Whole proteins extracted from the cultured adventitious roots of P. ginseng were separated by two-dimensional polyacrylamide gel electrophoresis (2-DE). Among the 1000 spots which were detected by silver staining, 113 spots were labeled and identified by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF MS). Our results showed that 40 proteins were identified among the 113 spots, with a hit ratio of 35.3%. A number of proteins identified on the 2-DE gels (30%; 16 spots) were involved in energy metabolism. These proteomic data will be helpful to better understand the physiological and pharmacological effects of P. ginseng.

• 한국임상수의학회지
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• 제19권1호
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• pp.49-54
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• 2002

Korean safflower (Carthami Flos) seed has been known to have healing effects on both bone fracture and osteoporosis. On the base of such a notice, this experiment was carried out to explore the effects of safflower seed on bone formation and bone repair. In addition, the healing mechanism was evaluated by analysing serum after feeding the seed to experimental. animals. The effect of Korean safflower seed were evaluated with 40 rats,3-month old. Forty Sprague-Dawley rats composed of 20 male and 20 female were underwent unilateral tibial defect and then fastened with unilateral fixators. The operated rats were divided into two groups depending on the composition of diet, such as positive control group fed normal diet (C-OP group) and safflower seed group fed 30% of safflower seed diet and 70% of normal diet (S-OP group). Postoperative radiographys were taken once in 2 weeks to evaluate callus formation for operated groups. In addition, a possible protein spots involved in bone recovery were examined using 2-Dimensional Gel Electrophoresis (2-DE). The comparison of the radiography between C-OP and S-OP group were showed that the safflower seed diet appeared to stimulate the formation of callus in the rat. On the images of 2-BE, it was able to identify possible five protein spots, having pl from 4 to 5 and molecular weight range from 24 to 26 kDa, involved in bone formation and repair, since no differing protein spots were found the two between groups except the five spots. No differences were observed between two groups before operation, but clear and bigger protein spots were observed from the S-OP group compared with C-OP group on 6 and 9 weeks post operation. These protein spots were, however, showed similar sizes and densities between two groups in 12 weeks later. The transformation of protein spots was suggested that these protein spots were involved in bone formation and recovery, in addition safflower seed might induce the formation of factors and activate these factors. In conclusion, this study suggest that safflower seed influence a variety of factors in the course of bone formation or the periods of remedy.

• Asian-Australasian Journal of Animal Sciences
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• 제29권9호
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• pp.1239-1246
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• 2016

The breeding of yaks is highly seasonal, there are many crucial proteins involved in the reproduction control program, especially in follicular development. In order to isolate differential proteins between mature and immature follicular fluid (FF) of yak, the FF from yak follicles with different sizes were sampled respectively, and two-dimensional gel electrophoresis (2-DE) of the proteins was carried out. After silver staining, the Image Master 2D platinum software was used for protein analysis and matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) was performed for differential protein identification. The expression level of transferrin and enolase superfamily member 1 (ENOSF1) was determined by Western blotting for verification analysis. The results showed that 2-DE obtained an electrophoresis map of proteins from mature and immature yak FF with high resolution and repeatability. A comparison of protein profiles identified 12 differently expressed proteins, out of which 10 of them were upregulated while 2 were downregulated. Western blotting showed that the expression of transferrin and ENOSF1 was enhanced with follicular development. Both the obtained protein profiles and the differently expressed proteins identified in this study provided experimental data related to follicular development during yak breeding seasons. This study also laid the foundation for understanding the microenvironment during oocyte development.

• Journal of Microbiology and Biotechnology
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• 제19권10호
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• pp.1109-1121
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• 2009

In an attempt to discover novel biomarker proteins in type 2 diabetes prognosis, we investigated the influence of hypoglycemic extracellular polysaccharides (EPS) obtained from the macrofungus Tremella fuciformis on the differential levels of plasma proteins in ob/ob mice using two-dimensional gel electrophoresis (2-DE). The 2-DE analysis demonstrated that 92 spots from about 900 visualized spots were differentially regulated, of which 40 spots were identified as principal diabetes-associated proteins. By comparing control with EPS-fed mice, we found that at least six proteins were significantly altered in ob/ob mice, including Apo A-I, IV, C-III, E, retinol-binding protein 4, and transferrin, and their levels were interestingly normalized after EPS treatment. Western blot analysis revealed that the altered levels of the two regulatory molecules highlighted in diabetes and obesity (e.g., resistin and adiponectin) were also normalized in response to EPS. The Mouse Diabetes PCR Array profiles showed that the expression of 84 genes related to the onset, development, and progression of diabetes were significantly downregulated in liver, adipocyte, and muscle of ob/ob mice. EPS might act as a potent regulator of gene expression for a wide variety of genes in ob/ob mice, particularly in obesity, insulin resistance, and complications from diabetes mellitus.

• 정보처리학회논문지D
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• 제15D권5호
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• pp.681-690
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• 2008

단백질체학에서 특정 조건 하에서 단백질의 기능 이상 및 구조 변형 유무를 규명하고 질병 과정을 추적하는 것은 중요한 연구이다. 일반적으로 단백질의 발현량 변화 분석에는 통계적 방법이 많이 사용되고 있으며 단백질 상용 이미지 분석 소프트웨어에서 제공하는 그래픽을 이용한 방법들도 있으나, 이 방법들은 많은 조직 내에 존재하는 수많은 단백질을 수동으로 비교해야 하는 어려움이 있다. 본 논문에서는 데이터베이스와 데이터마이닝 기법을 이용하여 OLAP 데이터 큐브와 Discovery-driven 탐색의 응용 방법을 제안한다. 데이터 큐브의 특성을 이용함에 의해서, 질병에 의해 발현량이 변하는 단백질 뿐 아니라 임상적 특성과 단백질의 영향 관계를 분석하는 것이 가능하다. 데이터 큐브에서 단백질의 발현량 변화 분석에 적합한 데이터 큐브의 척도와Discovery-driven 탐색을 위한 예외 지표를 제안하고, 특히 In-exception을 계산하는데 있어서의 계산량 감소 방안을 제시한다. 실험을 통해 폐암 2-DE 데이터에서 데이터 큐브와 Discovery-driven 방법이 유용함을 보인다.

• Molecules and Cells
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• 제24권2호
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• pp.268-275
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• 2007

Protein phosphorylation plays a key role in signal transduction in cells. Since phosphoproteins are present in low abundance, enrichment methods are required for their purification and analysis. Chemical derivatization strategies have been devised for enriching phosphoproteins and phosphopeptides. In this report, we employed a strategy that replaces the phosphate moieties on serine and threonine residues with a biotin-containing tag via a series of chemical reactions. Ribulose 1,5-bisphosphate carboxylase/oxygenase (RUBISCO)-depleted protein extracts prepared from Arabidopsis seedlings were chemically modified for 'biotin-tagging'. The biotinylated (previously phosphorylated) proteins were then selectively isolated by avidin-biotin affinity chromatography, followed by two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser-desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). This led to the identification of 31 protein spots, representing 18 different proteins, which are implicated in a variety of cellular processes. Despite its current technical limitations, with further improvements in tools and techniques this strategy may be developed into a useful approach.

• 한국수정란이식학회지
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• 제25권4호
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• pp.281-286
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• 2010

This study was to evaluate the protein profile of seminal plasma using 2-DE in Hanwoo. Seminal plasma was harvested from five mature Hanwoo, and seminal plasma protein was extracted by M-PER Mammalian Protein Extraction Reagent. Proteins were refined by clean-up kit and quantified by Bradford method until total protein was $300\;{\mu}l$. Immobilized pH gradient (IPG) strip was used 18 cm and 3~11 NL. SDS-PAGE was used 12% acrylamide gel. Each gels were visualized by comassie brilliant blue and silver staining. These spots were analyzed by MALDI-TOF MS and searched on NCBInr. The result, 20 proteins of 36 protein spots were searched through peptide sequencing on the NCBInr. 8 proteins profiled by 2-DE were proved through previous bovine studies and the name of each protein was albumin, nucleobindin, clusterin, TIMP-2, spermadhesin Z13, spermadhesin-1 and BSP proteins (BSP 30 kDa and BSP A1/A2). 12 new proteins were ATP synthase, protein MAK16 homolog, Transmembrane protein 214, E3 ubiquitin-protein ligase BRE1A, dual serine/threonine and tyrosine protein kinase, tissue factor pathway inhibitor 2, alpha-actinin-4, RUN domain-containing protein 3B, catenin alpha-1, protein-glutamine gamma-glutamyltransferase 2, plakophilin-1 and inter-alpha-trypsin inhibitor heavy chain H1 has not been previously described in the bovine seminal plasma study. These proteins may be contribute to define the type of proteins affecting fertility of male and improve the fertilizing ability of semen in Hanwoo.

• BMB Reports
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• 제40권1호
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• pp.72-81
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• 2007

In order to screen the aging related proteins in human normal colon epithelia, the comparative proteomics analysis was applied to get the two-dimensional electrophoresis (2-DE) profiles with high resolution and reproducibility from normal colon epithelial tissues of young and aged people. Differential proteins between the colon epithelia of two age groups were found with PDQuest software. The thirty five differential protein-spots were identified by peptide mass fingerprint (PMF) based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) and database searching. Among them there are sixteen proteins which are significantly up-regulated in the colonic mucosal epithelia of young people group, which include ATP synthase beta chain, electron transfer flavoprotein alpha-subunit, catalase, glutathione peroxidase 1, annexin A2 and heat shock cognate 71 kDa protein, etc.; There are nineteen proteins which are significantly up-regulated in the colonic mucosal epithelia of aged people group, which include far upstream element-binding protein 1, nucleoside diphosphate kinase B, protein disulfide-isomerase precursor and VDAC-2, etc.. The identified differential proteins appear to be involved in metabolism, energy generation, chaperone, antioxidation, signal transduction, protein folding and apoptosis. The data will help to understand the molecular mechanisms of human colon epithelial aging.

• Journal of Microbiology and Biotechnology
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• 제18권8호
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• pp.1364-1367
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• 2008

Oxidative stress damages all cellular constituents, and therefore, cell has to possess various defense mechanisms to cope. Saccharomyces cerevisiae, widely used as a model organism for studying cellular responses to oxidative stress, contains three glutathione peroxidase (Gpx) proteins. Among them, Gpx3 plays a major defense role against oxidative stress in S. cerevisiae. In this study, in order to identity the new interaction proteins of Gpx3, we carried out two-dimensional gel electrophoresis after immunoprecipitation (IP-2DE), and MALDI-TOF mass spectrometry. The results showed that several proteins including protein disulfide isomerase, glutaredoxin 2, and SSY protein 3 specifically interact with Gpx3. These findings led us to suggest the possibility that Gpx3, known as a redox sensor and ROS scavenger, has another functional role by interacting with several proteins with various cellular functions.

• 약학회지
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• 제54권3호
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• pp.180-186
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• 2010

Aldosterone, mineralocorticoid hormone, has important functions related to the regulation of blood pressure and balance of fluids and electrolytes in the distal region of the nephron. By genomic and non-genomic action of aldosterone, the physiological kidney functions are modulated. However, many of them except several kind of sodium channel have not been identified and analyzed yet. In this study, proteomic technologies with two-dimensional gel electrophoresis (2-DE) gel using aldosterone rat model were applied to analyze and identify the aldosterone dependently expressed proteins in rat kidney cortex. As a result, the established aldosterone rat model exhibited the normal physiological responses to aldosterone and modulated proteins were identified, which included 15 increased and 3 decreased proteins on 2-DE analysis. Among them, 11 proteins were identified as changed proteins by LC-MS/MS analysis. These proteins identified as aldosterone induced proteins were involved in several cellular pathways such as cytoskeleton remodeling, energy metabolism, amino acid metabolism, and chaperone process. In conclusion, our data could provide the insights into the new mechanism underlying regulation of kidney functions by aldosterone.