• 한국작물학회:학술대회논문집
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• 한국작물학회 2003년도 춘계 학술대회지
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• pp.51-51
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• 2003

• BMB Reports
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• 제40권6호
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• pp.853-860
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• 2007

Resistance to anticancer drugs is a major obstacle in the effective treatment of tumors. To understand the mechanisms responsible for multidrug resistance (MDR), a proteomic approach was used to identify proteins that were expressed in different levels by the adriamycinresistant human gastric cancer cell line, SGC7901/ADR, and its parental cell line, SGC7901. Two-dimensional gel electrophoresis (2-DE) and image analysis was used to determine which protein spots were expressed in different levels by the two cell lines. These spots were then partially identified using ESI-Q-TOF mass spectrometry, and the differential expressional levels of the partially identified proteins were then determined by western blot analysis and real-time RT-PCR. Additionally, the association of Nucleophosmin (NPM1), a protein that was highly expressed by SGC7901/ADR, with MDR was analyzed using siRNA. As a result of this study, well-resolved, reproducible 2-DE patterns of SGC7901/ADR and SGC7901 were established, and 16 proteins that may playa role in the development of thermo resistance were identified. Additionally, suppression of NPMl expression was found to enhance adriamycin chemosensitivity in SGC7901/ADR. These results provide a fundamental basis for the elucidation of the molecular mechanism of MDR, which may assist in the treatment of gastric cancer.

• Bulletin of the Korean Chemical Society
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• 제28권9호
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• pp.1499-1509
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• 2007

Proteomic analyses of synaptic vesicle fraction from rat brain have been performed for the better understanding of vesicle regulation and signal transmission. Two different approaches were applied to identify proteins in synaptic vesicle fraction. First, the isolated synaptic vesicle proteins were treated with trypsin, and the resulting peptides were analyzed using a high-pressure capillary reversed phase liquid chromatography/tandem mass spectrometry (cRPLC/MS/MS). Alternatively, proteins were separated by two-dimensional gel electrophoresis (2DE) and identified by matrix-assisted laser desorption ionization mass spectrometry (MALDI-TOF/MS). Total 18 and 52 proteins were identified from cRPLC/MS-MS and 2DE-MALDI-TOF-MS analysis, respectively. Among them only 2 proteins were identified by both methods. Of the proteins identified, 70% were soluble proteins and 30% were membrane proteins. They were categorized by their functions in vesicle trafficking and biogenesis, energy metabolism, signal transduction, transport and unknown functions. Among them, 27 proteins were not previously reported as synaptic proteins. The cellular functions of unknown proteins were estimated from the analysis of domain structure, expression profile and predicted interaction partners.

• Journal of Plant Biotechnology
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• 제39권1호
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• pp.81-92
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• 2012

단백질체학에서 이차원 전기영동 (2-DGE)은 고해상도 단백질 분리가 가능한 중요 기술 중 하나이다. 본 논문에서는 2-DGE 이미지 분석 일치도를 향상시킴으로써 작물 단백질 검출과 정량분석이 가능하도록 하는 전반적인 주요 장비, 시약 등을 자세히 기술한 프로토콜을 제시하고 있다. 이 프로토콜은 식물의 발달, 생물학적, 비생물학적 스트레스 반응 등과 관련된 작물 단백질 바이오마커를 개발하기 위한 목적에서 2-DGE를 처음 시도하는 연구자들에게 유용할 것으로 기대한다.

• 한국응용곤충학회지
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• 제29권3호
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• pp.157-164
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• 1990

한국산 멧누에(Bomdyx mandarina)chorion 유전자의 형질 발현기구를 규명하기 위한 연구의 일환으로 본 실험을 실시했다. 멧누에의 난각구조를 주사형 전자현마경에 의해 관찰한 결과 매우 특이적인 구조가 인정되었다. 즉 원추상의 불규칙 돌기에 의한 돌기구조층과 이 돌기 구조층을 덮고 있는 한 층의 얇은 덮개 구조층이 그것이다. 2차원 전기영동법에 의해 chorion 단백질을 분석한 결과, 난각을 구성하는 주요 단백질 성분은 등전점 4~6, 분자량 6~30 kd로 밝혀졌다. 특히 특이난각구조와 관련된 특이단백질 성분을 검출하였으며 이들의 대부분은 고cysteine 단백질인 것으로 추정했다. 이상의 연구결과에 의해 멧누에 난각의 특이 구조 형성에 따른 유전자발현기구 규명을 위한 기초자료가 얻어졌다.

• BMB Reports
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• 제39권6호
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• pp.703-708
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• 2006

During infection, the common respiratory tract pathogen Streptococcus pneumoniae encounters several environmental conditions, such as upper respiratory tract, lung tissue, and blood stream, etc. In this study, we examined the effects of blood on S. pneumoniae protein expression using a combination of highly sensitive 2-dimensional electrophoresis (DE) and MALDI-TOF MS and/or LC/ESI-MS/MS. A comparison of expression profiles between the growth in THY medium and THY supplemented with blood allowed us to identify 7 spots, which increased or decreased two times or more compared with the control group: tyrosyl-tRNA synthetase, lactate oxidase, glutamyl-aminopeptidase, L-lactate dehydrogenase, cysteine synthase, ribose-phosphate pyrophosphokinase, and orotate phosphoribosyltransferase. This global approach can provide a better understanding of S. pneumoniae adaptation to its human host and a clue for its pathogenicity.

• 방사선산업학회지
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• 제5권1호
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• pp.1-5
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• 2011

Since proteomics can be employed to compare changes in the expression levels of many proteins under particular genetic and environmental conditions, using mass spectrometry to establish radiation stimulon, we performed two-dimensional gel electrophoresis and identified E. coli proteins whose expressions are affected by high dose of ionizing radiation. After exposure to 3 kGy, it was found that 6 proteins involved in carbon and energy metabolism were reduced. Although 4 of 7 protein spots showing a significant increase in expression level were neither identified nor classified, uridine phosphorylase (Udp), superoxide dismutase (SodB), and thioredoxin-dependent thiol peroxidase (Bcp) were proven to be up-regulated after irradiation. This suggests that E. coli subjected to high doses of radiation (3 kGy) may operate a defense system that is able to detoxify reactive oxygen species and stimulate the salvage pathway of nucleotide synthesis to replenish damaged DNA.

• 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.

• Journal of Microbiology and Biotechnology
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• 제21권2호
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• pp.136-146
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• 2011

This study provides first-hand proteomic data on the survival strategy of Anabaena sp. PCC 7120 when subjected to long-term iron-starvation conditions. 2D-gel electrophoresis followed by MALDI-TOF/MS analysis of iron-deficient Anabaena revealed significant and reproducible alterations in ten proteins, of which six are associated with photosynthesis and respiration, three with the antioxidative defense system, and the last, hypothetical protein all1861, conceivably connected with iron homeostasis. Iron-starved Anabaena registered a reduction in growth, photosynthetic pigments, PSI, PSII, whole-chain electron transport, carbon and nitrogen fixation, and ATP and NADPH content. The kinetics of hypothetical protein all1861 expression, with no change in expression until day 3, maximum expression on the $7^{th}$ day, and a decline in expression from the $15^{th}$ day onward, coupled with in silico analysis, suggested its role in iron sequestration and homeostasis. Interestingly, the up-regulated FBP-aldolase, Mn/Fe-SOD, and all1861 all appear to assist the survival of Anabeana subjected to iron-starvation conditions. Furthermore, the $N_2$-fixation capabilities of the iron-starved Anabaena encourage us to recommend its application as a biofertilizer, particularly in iron-limited paddy soils.

• Asian Pacific Journal of Cancer Prevention
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• 제14권1호
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• pp.367-372
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• 2013

Effects of the Epstein-Barr virus (EBV) on cellular protein expression are essential for viral pathogenesis. To characterize the cellular response to EBV infection, differential proteomes of gastric epithelial AGS cells were analyzed with two-dimensional gel electrophoresis (2-DE) followed by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) and liquid chromatography electrospray/ionization ion trap (LC-ESI-IT) mass spectrometry identification. Mass spectrometry identified 9 altered cellular proteins, including 5 up-regulated and 4 down-regulated proteins after EBV infection. Notably 2-DE analysis revealed that EBV infection induced increased expression of heat shock cognate 71 kDa protein, actin cytoplasmic 1, pyridoxine-5'-phosphate oxidase, caspase 9, and t-complex protein 1 subunit alpha. In addition, EBV infection considerably suppressed those cellular proteins of zinc finger protein 2, cyclin-dependent kinase 2, macrophage-capping protein, and growth/differentiation factor 11. Furthermore, the differential expressional levels of partial proteins (cyclin-dependent kinase 2 and caspase 9) were confirmed by Western blot analysis.Thus, this work effectively provided useful protein-related information to facilitate further investigation of the mechanisms underlying EBV infection and pathogenesis.