• Title/Summary/Keyword: Proteome analysis

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Improved Viability and Proteome Analysis of Lactobacillus fermentum KLB12 upon Heat Stress (Lactobacillus fermentum KLB12의 열 전처리에 따른 열 스트레스 내성 증진 및 프로테옴 변화)

  • 김주현;박미영;김승철;윤현식;소재성
    • KSBB Journal
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    • v.18 no.4
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    • pp.294-300
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    • 2003
  • In the previous study, we have isolated several vaginal lactobacilli from Korean woman and selected one of them (KLB12) for further study, which was indentified as Lactobacillus fermentum by sequence analysis of 16S rRNA gene. Formulated L. fermentum KLB12 can be used for ecological treatment of bacterial vaginosis. For pharmaceutical formulation, the spray-drying process is required where stress such as high temperature is routinely applied. In this study, we found that heat stress at 60$^{\circ}C$ for 20∼30min reduced the viable cell population of KLB12 by 10$\sub$6/~10$\sub$9/. However, adaptation of KLB12 cells at 52$^{\circ}C$ made them more thermotolerant upon exposure to 60$^{\circ}C$. The level of thermal protection in RSM (reconstituted skim milk) by adaptation in acid (pH 5), cold (4$^{\circ}C$), ethanol (3%), NaCI (0.3M) was also examined. Although not as efficient as the homologous stress, adaptations in both cold and NaCI gave considerable cross protection against heat stress. When chloramphenicol was added during heat adaptation, adaptation effect was abolished. This suggests that de novo protein synthesis is necessary during the adaptation process. Important changes in proteome during heat adaptation was examined with two-dimensional gel electrophoresis.

MicroRNA super-resolution imaging in blood for Alzheimer's disease

  • Mirae Lee;Jiwon Woo;Sang Tae Kim;Minho Moon;Sang Yun Kim;Hanna Cho;Sujin Kim;Han-Kyeol Kim;Jeong-Yoon Park
    • BMB Reports
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    • v.56 no.3
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    • pp.190-195
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    • 2023
  • We propose a novel blood biomarker detection method that uses miRNA super-resolution imaging to enable the early diagnosis of Alzheimer's disease (AD). Here, we report a single-molecule detection method for visualizing disease-specific miRNA in tissue from an AD mice model, and peripheral blood mononuclear cells (PBMCs) from AD patients. Using optimized Magnified Analysis of Proteome (MAPs), we confirmed that five miRNAs contribute to neurodegenerative disease in the brain hippocampi of 5XFAD and wild-type mice. We also assessed PBMCs isolated from the whole blood of AD patients and a healthy control group, and subsequently analyzed those samples using miRNA super-resolution imaging. We detected more miR-200a-3p expression in the cornu ammonis 1 and dentate gyrus regions of 3 month-old 5XFAD mice than in wild-type mice. Additionally, miRNA super-resolution imaging of blood provides AD diagnosis platform for studying miRNA regulation inside cells at the single molecule level. Our results present a potential liquid biopsy method that could improve the diagnosis of early stage AD and other diseases.

An integrated bioinformatics analysis of mouse testis protein profiles with new understanding

  • Liu, Fujun;Wang, Haiyan;Li, Jianyuan
    • BMB Reports
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    • v.44 no.5
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    • pp.347-351
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    • 2011
  • The testis is major male gonad responsible for spermatogenesis and steroidogenesis. Much knowledge is still remained to be learned about the control of these events. In this study, we performed a comprehensive bioinformatics analysis on 1,196 mouse testis proteins screened from public protein database. Integrated function and pathway analysis were performed through Database for Annotation, Visualization and Integrated Discovery (DAVID) and ingenuity Pathway Analysis (IPA), and significant features were clustered. Protein membrane organization and gene density on chromosomes were analyzed and discussed. The enriched bioinformatics analysis could provide clues and basis to the development of diagnostic markers and therapeutic targets for infertility and male contraception.

Effects of the Applications of Clay Minerals on the Early Growth of Red Pepper in Growing Medium (점토광물 처리에 따른 상토에서 고추의 초기생장 효과)

  • Lee, Dong-Gi;Lee, Seok-Eon;Kim, Deok-Hyun;Hong, Hyeon-Ki;Nam, Ju-Hyun;Choi, Jong-Soon;Lee, Moon-Soon;Woo, Sun-Hee;Chung, Keun-Yook
    • Horticultural Science & Technology
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    • v.30 no.4
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    • pp.463-470
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    • 2012
  • As the advanced seeding technology through use of plug tray for good cultivation of seeds was propagated along with the expansion and development of horticultural industry, the use of bed soils as growing medium has recently been increased. In this study, the effects of the four clay minerals such as illite, phyllite, zeolite, and bentonite on the early growth of red pepper in the bed soil were investigated. Furthermore, proteome analysis for the leaf and stem samples of red pepper treated with only illite was performed. Of the seedling cultured, the healthy and regular size seeds were selected and cultivated in the pots, after they were treated with four clay minerals. The experiment was performed during the whole six weeks in the glasshouse of the Chungbuk National University. The growth lengths, fresh and dry weights of red pepper were significantly higher in the treatments of illite, phyllite, zeolite, and bentonite than in the control. In addition, the uptake of $K^+$, $Ca^{2+}$, and $Mg^{2+}$ were higher in the treatment of illite, phyllite, zeolite, and bentonite than in the control. The 2-DE patterns for the red pepper by the applications of illite, phyllite, zeolite, and bentonite were similar to each other. Therefore, compared to the samples of control, the proteome analysis for the samples of red pepper treated by only illite were performed. Proteome analysis for red pepper showed that plastid fructose-1, 6-bisphosphate aldolase class 1, aldolase, and glyceraldehydes 3-phosphate dehydrogenase, all of which were involved in the energy metabolism, were highly expressed in leaf tissue by illite treatment. In stem tissue, NAD-dependent formate dehydrogenase involved in energy metabolism, potassium transport protein, and GIA/RGA-like gibberellins response modulator were highly expressed. Based on the results obtained from the proteome analysis, it appears that the proteins specifically and differentially expressed on the illite treatment may be involved in the enhanced growth of red pepper. The identification of some proteins involved in the response of vegetable crops to the treatment of clay mineral can provide new insights that can lead to a better elucidation and understanding of mechanism on their molecular basis.

Analysis of Fatty Acyl Groups of Diacyl Galactolipid Molecular Species by HPLC/ESI-MS with In-source Fragmentation

  • Gil, Ji-Hye;Hong, Jong-Ki;Choe, Joong-Chul;Kim, Young-Hwan
    • Bulletin of the Korean Chemical Society
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    • v.24 no.8
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    • pp.1163-1168
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    • 2003
  • The structures of molecular species of galactolipids, such as monogalactosyl diacylglycerol (MGDG) and digalactosyl diacylglycerol (DGDG), isolated from wheat flour have been investigated using negative-ion electrospray ionization (ESI) mass spectrometry interfaced with high performance liquid chromatography (HPLC). According to the result of HPLC analysis, MGDG and DGDG were found to consist of mixtures of five and four molecular species, respectively. The galactolipids have been also analyzed to determine their fatty acid compositions, using HPLC/ESI-MS combined with in-source (or cone voltage) fragmentation. HPLC/ ESI-MS is very useful for one-step analysis of mixtures of galactolipids with a small sample quantity. Especially, the carboxylate anions produced in in-source fragmentations of the negative-ion of each component separated by HPLC provide valuable information on the composition of its fatty acyl chains.

Microbial Diversity Information Facility: Bacteriology Insight Orienting System (BIOS)

  • Shimura, Junko;Shimiz, Hideyukiu;Tsuruwaka, Keiji;Moritani, Yukimitsu;Miyazaki, Kenji;Tsugita, Akira;Watanabe, Makoto M.
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.04a
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    • pp.135-141
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    • 2000
  • Global Biodiversity is common interest of humans for better health and sustainable development of the society. To provide access and analysis on microbial diversity information, Bacteriology Insight Orienting System (BIOS) has been developed. BIOS contains 6402 species and subspecies names of bacteria and archaea, 2606 names of cyanobacteria by March 2000. BIOS of which web based analytical tool provides windows to compare the results of phylogenetic analysis based on 16S rDNA sequence and the results of cluster analysis on proteome profiling. The sequence data and 2 dimensional gel electrophoresis analysis data were accumulated in BIOS database content for cyanobacteria reclassification and taxonomy. (BIOS URL: http.://www-sp2000ao.nies.go.jp/bios/index.html).

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Informatics for protein identification by tandem mass spectrometry; Focused on two most-widely applied algorithms, Mascot and SEQUEST

  • Sohn, Chang-Ho;Jung, Jin-Woo;Kang, Gum-Yong;Kim, Kwang-Pyo
    • Bioinformatics and Biosystems
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    • v.1 no.2
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    • pp.89-94
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    • 2006
  • Mass spectrometry (MS) is widely applied for high throughput proteomics analysis. When large-scale proteome analysis experiments are performed, it generates massive amount of data. To search these proteomics data against protein databases, fully automated database search algorithms, such as Mascot and SEQUEST are routinely employed. At present, it is critical to reduce false positives and false negatives during such analysis. In this review we have focused on aspects of automated protein identification using tandem mass spectrometry (MS/MS) spectra and validation of the protein identifications of two most common automated protein identification algorithms Mascot and SEQUEST.

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Characterization of a PyrR-deficient Mutant of Bacillus subtilis by a Proteomic Approach (프로테옴 분석에 의한 Bacillus subtilis PyrR 돌연변이체의 특성)

  • Seul, Keyung-Jo;Cho, Hyun-Soo;Ghim, Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.39 no.1
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    • pp.9-19
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    • 2011
  • The Bacillus subtilis pyrimidine biosynthetic (pyr) operon encodes all of the enzymes for the de novo biosynthesis of Uridine monophosphate (UMP) and additional cistrones encoding a uracil permease and the regulatory protein PyrR. The PyrR is a bifunctional protein with pyr mRNA-binding regulatory funtion and uracil phosphoribosyltransferase activity. To study the global regulation by the pyrR deletion, the proteome comparison between Bacillus subtilis DB104 and Bacillus subtilis DB104 ${\Delta}$pyrR in the minimal medium without pyrimidines was employed. Proteome analysis of the cytosolic proteins from both strains by 2D-gel electrophoresis showed the variations in levels of protein expression. On the silver stained 2D-gel with an isoelectric point (pI) between 4 and 10, about 1,300 spots were detected and 172 spots showed quantitative variations in which 42 high quantitatively variant proteins were identified. The results showed that production of the pyrimidine biosynthetic enzymes (PyrAA, PyrAB, PyrB, PyrC, PyrD, and PyrF) were significantly increased in B. subtilis DB104 ${\Delta}$pyrR. Besides, proteins associated carbohydrate metabolism, elongation protein synthesis, metabolism of cofactors and vitamins, motility, tRNA synthetase, catalase, ATP-binding protein, and cell division protein FtsZ were overproduced in the PyrR-deficient mutant. Based on analytic results, the PyrR might be involved a number of other metabolisms or various phenomena in the bacterial cell besides the pyrimidine biosynthesis.

Proteome Analysis of Chicken Embryonic Gonads: Identification of Major Proteins from Cultured Gonadal Primordial Germ Cells

  • Lee, Sang-In;Han, Beom-Ku;Park, Sang-Hyun;Kim, Tae-Min;Sin, Sang-Soo;Lee, Young-Mok;Kim, Hee-Bal;Lim, Jeong-Mook;Han, Jae-Yong
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2005.11a
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    • pp.66-67
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    • 2005
  • The domestic chicken (Gallus gallus) is an important model for research in developmental biology because its embryonic development occurs in ovo. To examine the mechanism of embryonic germ cell development, we constructed proteome map of gonadal primordial germ cells (gPGC) from chicken embryonic gonads. Embryonic gonads were collected from 500 embryos at 6 day of incubation, and the gPGC were cultured in vitro until colony formed. After 7-10 days in cultured gPGC colonies were separated from gonadal stroma cells (GSCs). Soluble extracts of cultured gPGCs were then fractionated by two-dimensional gel electrophoresis (pH 4-7). A number of protein spots, including those that displayed significant expression levels, were then identified by use of matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry and LC-MS/MS. Of the 89 gPGC spots examined, 50 yielded mass spectra that matched avian proteins found in on-line databases. Proteome map of thistype will serve as an important reference for germ cell biology and transgenic research.

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Differential Proteome Expression of in vitro Proliferating Hanwoo Stromal Vascular Cells from Omental, Subcutaneous and Intramuscular Depots in Response to Hormone Deprivation and IGF-1, Estradiol-17β Addition

  • Rajesh, Ramanna Valmiki;Kim, Seong-Kon;Park, Min-Ah;Kwon, Seulemina;Chang, Jong-Soo;Yoon, Du-Hak;Kim, Tae-Hun;Lee, Hyun-Jeong
    • Journal of Animal Science and Technology
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    • v.52 no.3
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    • pp.175-186
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    • 2010
  • The aim of this study was to analyze the proteome expressions of proliferating stromal vascular cells from Hanwoo omental, subcutaneous and intramuscular depots subjected to hormone deprivation and IGF-1, Estradiol-$17{\beta}$ addition. For hormone deprivation or addition studies, the cells were either grown in 10% charcoal-dextran stripped fetal bovine serum (CD-FBS) or in 10% FBS supplemented medium. Further, to analyze the effect of insulin like growth factor (IGF-1) and $17\beta$-Estradiol (E2), cells were grown in 10% CD-FBS containing IGF-1 (10 ng/ml) or E2 (10 nM). The results showed that hormone deprivation had a negative impact on proliferation among the cells from all depots without any growth difference. On comparison of proliferation levels, higher levels were observed in cells that were grown in 10% FBS than in 10% CD-FBS alone or with IGF-1/E2. Proteome expression from preadipocytes grown in hormone deprivation conditions were compared by 2D-DIGE and MALDIToF/ToF. A total of twelve different proteins were found to be differentially expressed under hormone deprivation conditions. Further, our proteomic analysis with DIGE under IGF-1 and E2 addition revealed four proteins with differential expression levels. Moreover, the results highlighted in this study offer a role for each differentially expressed protein with respect to their effect in positive or negative regulation on proliferation.