• 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.

• Animal cells and systems
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• 제4권2호
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• pp.145-150
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• 2000

Retinoic acid (RA) evokes pattern duplication in the regenerating salamander limb. Interestingly, it also enhances dedifferentiation in the regenerate by the morphological, histological and biochemical criteria. To examine whether there is any correlation between the RA-evoked pattern duplication and de novo protein synthetic profile in the regenerating salamander limb, especially during dedifferentiation, we analyzed stage-specific protein synthesis pattern in the normal and RA-treated regenerating limbs by metabolic labeling followed by two-dimensional gel electrophoresis. In the regenerating limbs without RA treatment, a few hundred kinds of proteins were found to be synthesized at the stage of wound healing and the total number of protein synthesized increased greatly as regeneration proceeded. The same trend was also observed in the RA-treated regenerating limbs. Interestingly, some protein spots were noted to be either newly synthesized or highly expressed by the RA treatment especially at the stage of dedifferentiation. The results shows that the enhancement of dedifferentiation state after the RA treatment correlates well with the protein synthesis profile, and suggest that those proteins are important for the RA-evoked pattern duplication in the regenerating limbs of salamander.

• 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.

• The Korean Journal of Physiology and Pharmacology
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• 제8권4호
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• pp.207-211
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• 2004

The purpose of the present study was to evaluate the expression of cardiac marker protein in rabbit cardiac tissue that was exposed to ischemic preconditioning (IPC), or ischemiareperfusion injury (IR) using two-dimensional gel electrophoresis (2DE) and matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). We compared 2DE gels of control (uninjured) cardiac tissue with those of IPC and IR cardiac tissue. Expression of one protein was detected in IR heart tissue, however the protein was not detected in the samples of control and IPC tissue. To further characterize the detected protein molecule, the protein in the 2D gel was isolated and subjected to trypsin digestion, followed by MALDI-MS. The protein was identified as myoglobin, which was confirmed also by Western blot analysis. These results are consistent with previous studies of cardiac markers in ischemic hearts, indicating myoglobin as a suitable marker of myocardial injury. In addition, the present use of multiple techniques indicates that proteomic analysis is an appropriate means to identify cardiac markers in studies of IPC and IR.

• Journal of Animal Science and Technology
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• 제45권5호
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• pp.731-738
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• 2003

근육의 생화학적 특성을 단백질 수준에서 분석하기 위하여 3원교잡종 돼지 3개체를 선발하고, white muscle은 longissimus dorsi muscle을 red muscle은 soleus muslce을 분리하여 분석에 이용하였다. 각 근육조직은 수용성, 불수용성 단백질 및 총단백질로 분리하여 추출하였고, 이차원전기영동 분석을 위하여 17cm 길이의 immobilized pH gradient strip (Bio-Rad, 3-10NL)과 12% acrylamide gel을 이용하여 전개하였다. 각각의 gel은 coomassie stain과 silver stain을 통하여 가시화 하였고, PDQuest software을 통하여 단백질 발현양상을 분석하였다. 하나의 gel에서 평균 600개 이상의 단백질 spot을 관찰하였으며, 반복실험을 통하여 white muscle과 red muscle간에 발현의 차이를 보이는 5개의 단백질 spot을 확인할 수 있었다. 5개의 spot 중 4개의 단백질은 측정된 분자량과 pI값이 troponin I, T 및 myoglobin의 수치값과 유사한 것으로 확인되었다. 그러나, 1개의 spot은 오차범위 내에서 유사한 단백질을 확인할 수 없었다. 5개의 단백질 spot중 1개(spot 1)는 white muscle에서, 4개의 spot(spot 2～5)은 red muscle에서 높게 발현됨을 확인하였으며, 특히 spot 4의 경우 white muscle 보다 red muscle에서 평균 14.6배 높게 발현됨을 확인하였다. 본 연구는 근육의 생화학적 특성을 이해하는데 중요한 기초 자료로 활용될 수 있으며, 앞으로 white muscle과 red muscle의 단백질 발현 분석을 통하여 단백질 수준에서의 생화학적 특성에 관한 연구가 충분히 진행되어야 할 것이다.

• Journal of Microbiology and Biotechnology
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• 제20권3호
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• pp.558-562
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• 2010

SDS-PAGE of extracted surface-associated proteins of Lactobacillus rhamnosus strains E/N, Oxy, and Pen, was performed. The obtained protein patterns allowed differentiation of the examined strains, which was not accomplished by the commonly used RAPD genotypic method. The differentiation by the SDS-PAGE method proved to be a useful tool for strain-specific identification, which was further confirmed by 2DE analysis. Therefore, it can be used as an alternative or complementary method for both conventional and genotypic identification procedures, especially when closely related lactobacilli isolates are identified.

• 생명과학회지
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• 제13권3호
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• pp.248-251
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• 2003

갑상선호르몬의 전구체인 thyroglobulin (Tg)이 어떻게 dimerization이 일어나는지를 조사하였다, 변성 혹은 비변성전기영동법으로 관찰한 결과 새롭게 만들어지기 시작한 Tg는 15분에 처음으로 구조적으로 불안정하지만 folded monomer가 관찰되었다. 그 이후에 monomer가 dimer로 바뀌어 결국 30분에 dimer Tg가 만들어지기 시작하여 40분에는 dimer Tg가 관찰되었다. 최종적으로 60분에 완전하게 dimerization된 Tg가 만들어졌다.

• Food Science and Biotechnology
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• 제18권1호
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• pp.249-252
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• 2009

Two-dimensional gel electrophoresis (2-DE) was employed to assess the thermo-tolerance characteristics of Bifrdobacterium infantis ATCC 27920 to mild heat adaptation. When exposed to various heat levels, pH, and hydrogen peroxide ($H_2O_2$) stress conditions, B. infantis ATCC 27920 exhibited high level of stress resistance. Under mild-heat treatment ($46^{\circ}C$), no significant change in viability level was observed after 2 hr. Interestingly, improved viability was observed in mild-heat adapted ($46^{\circ}C$ for 1 hr) cultures exposed to $55^{\circ}C$, in comparison to control experiments. Viability was not affected by pH, bile, and $H_2O_2$ stress conditions. 2-DE analysis revealed those mild-heat adaptation up-regulated 4 proteins and down-regulated 3 proteins. Among these protein spots, isopropyhnalate dehydratase (leuD), glycosyltransferase (glgA), and ribosomal protein L5 (rp1E) were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALD1-TOF/MS).

• 한국임상수의학회지
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• 제38권1호
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• pp.21-26
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• 2021

Pyometra, a common disorder in intact bitches, can lead to canine sepsis. Identification of biomarkers for sources of infection in the uterus using two-dimensional electrophoresis (2-DE)-mass spectrometry (MS) analysis may enable the discovery of novel diagnostic markers of sepsis. Toward this end, surgically resected uterus samples from four bitches (three pyometra and one healthy) were randomly selected for 2-DE-MS, which identified 32 differentially expressed proteins, including seven inflammatory proteins, five non-inflammatory proteins, and 20 functionally unknown proteins. Despite the limited information on canine uterus proteomics, we suggest the potential use of differentially expressed uterus proteins as candidate biomarkers to discover targets to attenuate inflammation in pyometra. Further identification of the functionally unknown proteins is warranted.

• 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.