• Title/Summary/Keyword: 2-Dimensional Electrophoresis

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Genetic Variation of High Molecular Weight Glutenin (HMW-Glu) Subunit in Korean Wheat

  • Hong, Byung-Hee;Park, Chul-Soo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.43 no.4
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    • pp.259-263
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    • 1998
  • High molecular weight glutenin (HMW-Glu) subunit compositions of 73 Korean wheat cultivars and experimental lines were evaluated by using one dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This method is suitable for obtaining a good resolution of 1Dx2 and 1Ax2$^*$ without adverse effects on separation of other HMW-Glu subunits. Korean wheats examined in this study could be divided into 15 different groups on the basis of HMW-Glu subunit compositions. From the wheat lines tested, it was identified that there were three alleles at the Glu-Al, five at the Glu-Bl and three at the Glu-D1 loci. The null allele of the Glu-Al was occurred in high frequency (79.4%), while low frequencies for 1Ax1 (12.3%) and 1Ax2$^*$(8.2%) were found. High frequency (75.3%) of the subunit pairs of 1Bx7+1By8 at the Glu-Bl loci compared with other subunits was found. The frequencies of subunits 1Dx2. 2+1Dy12 and 1Dx2+1Dy12 from the Glu-D1 loci were 54. 8% and 37.0%, respectively. However, a few Korean wheat lines (8.2%) carried 1Dx5 + 1Dy10 subunit pair which are responsible for good breadmaking quality. The information of HMW-Glu subunit compositions provide a useful tool to characterize wheat lines, and can be directly used in selection of breeding lines of different end-use properties.

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TNF-${\alpha}$ Up-regulated the Expression of HuR, a Prognostic Marker for Ovarian Cancer and Hu Syndrome, in BJAB Cells

  • Lee, Kyung-Yeol
    • IMMUNE NETWORK
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    • v.4 no.3
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    • pp.184-189
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    • 2004
  • Background: Hu syndrome, a neurological disorder, is characterized by the remote effect of small cell lung cancer on the neural degeneration. The suspicious effectors for this disease are anti-Hu autoantibodies or Hu-related CD8+ T lymphocytes. Interestingly, the same effectors have been suggested to act against tumor growth and this phenomenon may represent natural tumor immunity. For these diagnostic and therapeutic reasons, the demand for antibodies against Hu protein is rapidly growing. Methods: Polyclonal and monoclonal antibodies were generated using recombinant HuR protein. Western blot analyses were performed to check the specificity of generated antibodies using various recombinant proteins and cell lysates. Extracellular stimuli for HuR expression had been searched and HuR-associated proteins were isolated from polysome lysates and then separated in a 2-dimensional gel. Results: Polyclonal and monoclonal antibodies against HuR protein were generated and these antibodies showed HuR specificity. Antibodies were also useful to detect and immunoprecipitate endogenous HuR protein in Jurkat and BJAB. This report also revealed that TNF-${\alpha}$ treatment in BJAB up-regulated HuR expression. Lastly, protein profile in HuR-associated mRNAprotein complexes was mapped by 2-dimensional gel electrophoresis. Conclusion: This study reported that new antibodies against HuR protein were successfully generated. Currently, project to develop a diagnostic kit is in process. Also, this report showed that TNF-${\alpha}$ up-regulated HuR expression in BJAB and protein profile associated with HuR protein was mapped.

Identification of the Proteins Expressed at Optimum Marbling in Hanwoo Loin (한우육의 육질에 중요 영향을 미치는 근내지방축적 최적기에 발현되는 단백질 동정)

  • 임진규
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.28 no.3
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    • pp.579-585
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    • 1999
  • Proteins from the loin tissues age ranged from 0 to 24 months of ten Korean cattle were extracted, separated and compared on two dimensional(2 D) gels to identify the proteins whose expression is highly correlated to marbling. We also compared the difference of loin proteins between castrated and non castrated bull cows on two dimensional gels. As the marbling in the loin of the cattle is optimized at 18 to 24 months, eight proteins expressed significantly higher level in 24 month than in 0 or 6 month were selected in terms of isoelectric points(pIs) and molecular weights. Using these values, we searched the Swiss Prot database via the ExPASy molecular biology server with TagIdent program. The proteins with the nearest molecular weights and isoelectric points were selected from the lists. These possible candidates were confirmed by N terminal microsequencing of the eight selected proteins. Three proteins, myoglobin, hemoglobin and ATPase, whose N termini were not blocked could be microsequenced and found to be exactly matched to the selected candidates. It is suggested that the proteins increasingly expressed in marbling periods can be involved in meat color, lipid transport and flavor improvement.

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Cloned Placenta of Korean Native Calves Died Suddenly at Two Months after Birth Displays Differential Protein Expression

  • Kim Hong Rye;Kang Jae Ku;Lee Hye Ran;Yoon Jong Taek;Seong Hwan Hoo;Jung Jin Kwan;Park Chang Sik;Jin Dong Il
    • Reproductive and Developmental Biology
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    • v.29 no.2
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    • pp.63-68
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    • 2005
  • Cloned calves derived from somatic cell nuclear transfer (SCNT) have been frequently lost by sudden death at 1 to 3 month following healthy birth. To address whether placental anomalies are responsible for the sudden death of cloned calves, we compared protein patterns of 2 placentae derived from SCNT of Korean Native calves died suddenly at two months after birth and those of 2 normal placentae obtained from AI fetuses. Placental proteins were separated using 2-Dimensional gel electrophoresis. Approximately 800 spots were detected in placental 2-D gel stained with coomassie-blue. Then, image analysis of Malanie III (Swiss Institute for Bioinformatics) was performed to detect variations in protein spots between normal and SCNT placentae. In the comparison of normal and SCNT samples, 8 spots were identified to be up-regulated proteins and 24 spots to be down-regulated proteins in SCNT placentae, among which proteins were high mobility group protein HMG1, apolipoprotein A-1 precursor, bactenecin 1, tropomyosin beta chain, $H^+-transporting$ ATPase, carbonic anhydrase II, peroxiredoxin 2, tyrosine-rich acidic matrix protein, serum albumin precursor and cathepsin D. These results suggested that the sudden death of cloned calves might be related to abnormal protein expression in placenta.

Genomic and Proteomic Profiling of the Cadmium Cytotoxic Response in Human Lung Epithelial Cells

  • Choi, Kwang-Man;Youn, Hyung-Sun;Lee, Mi-Young
    • Molecular & Cellular Toxicology
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    • v.5 no.3
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    • pp.198-206
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    • 2009
  • Microarray and proteomic expression patterns in response to cadmium exposure were analyzed in human lung epithelial cells. Among 35,000 genes analyzed by cDNA microarray, 228 genes were up-regulated and 99 genes were down-regulated, based on a fold change cut-off value of ${\geq}2$. Combining two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization-time-of-flight-mass spectrometry (MALDI-ToF-MS), 25 of 629 protein spots showed fold changes in expression ${\geq}2$ (17 up-regulated, 8 down-regulated). After comparing the cDNA microarray and proteomic analyses, only transglutaminase 2, translation elongation factor 1 alpha 1, and glyceraldehyde-3-phosphate dehydrogenase showed overlapping signals in the cDNA microarray and proteomic analyses, whereas the remaining differentially expressed proteins showed large discrepancies with respect to mRNA expression.

Disassembly of Chlorophyll-Protein Complexes in Arabidopsis thaliana during Dark-Induced Foliar Senescence

  • Lee, Choon-Hwan
    • Journal of Plant Biology
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    • v.39 no.4
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    • pp.301-307
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    • 1996
  • The disassembly of Chl-protein complexes during dark-induced senescence (DIS) was investigated using detached third and fourthleaves of 21$\pm$1 day-old Arabidopsis thaliana. Although Chl content decreased linearly after 1 d, a significant decrease of photochemical effeciency (Fv/Fm) was observed after 2 d. In experiments using native green gel electrophoresis of Chl-protein complexes combined with additional two-dimensional SDS-PAGE analysis, we could observe the degradation of both photosystems after 2 d. Although light-harvesting complex(LHC) for PSI (LHCI) was degraded first in PSI complex, small PSII apoproteins including CP47/CP43 and D1/D2 apoproteins were degraded first in PSII complexes. LHC for PSII (LHCII) trimers were stable until 4 d. The level of LHCII monomers was increased until 3 and decreased thereafter, resulting in the increase of free pigments. These results suggest that the disassembly process of PSI is different from that of PSII.

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Toxicoproteomics in the Study of Aromatic Hydrocarbon Toxicity

  • Cho, Chang-Won;Kim, Chan-Wha
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.3
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    • pp.187-198
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    • 2006
  • The aromatic hydrocarbons (AHs), which include benzene, polycyclic aromatic hydrocarbons, and dioxin, are important chemical and environmental contaminants in industry that usually cause various diseases. Over the years, numerous studies have described and evaluated the adverse health effects induced by AHs. Currently, "Omics" technologies, transcriptomics and proteomics, have been applied in AH toxicity studies. Proteomics has been used to identify molecular mechanisms and biomarkers associated with global chemical toxicity. It could enhance our ability to characterize chemical-induced toxicities and to identify noninvasive biomarkers. The proteomic approach (e.g. 2-dimensional electrophoresis [2-DE]), can be used to observe changes in protein expression during chemical exposure with high sensitivity and specificity. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) and electrospray ionization-quadrupole (ESI-Q)-TOF MS/MS are recognized as the most important protein identification tools. This review describes proteomic technologies and their application in the proteomic analysis of AH toxicity.

Isolation of the Arabidopsis Phosphoproteome Using a Biotin-tagging Approach

  • Kwon, Sun Jae;Choi, Eun Young;Seo, Jong Bok;Park, Ohkmae K.
    • Molecules and Cells
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    • v.24 no.2
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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.

Characterization of Resistive Switching in PVP GQD / HfOx Memristive Devices (PVP GQD / HfOx 구조를 갖는 전도성 필라멘트 기반의 저항성 스위칭 소자 특성)

  • Hwang, Sung Won
    • Journal of the Semiconductor & Display Technology
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    • v.20 no.1
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    • pp.113-117
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    • 2021
  • A composite active layer was designed based on graphene quantum dots, which is a low-dimensional structure, and a heterogeneous active layer of graphene quantum dots was applied to the interfacial defect structure to overcome the limitations. Increasing to 1.5~3.5 wt % PVP GQD, Vf changed from 2.16 ~ 2.72 V. When negative deflection is applied to the lower electrode, electrons travel through the HfOx/ITO interface. The Al + ions are reduced and the device dominates at low resistance. In addition, as the PVP GQD concentration increased, the depth of the interfacial defect decreased, and the repetition of appropriate electrical properties was confirmed through Al and HfOx/ITO. The low interfacial defects help electrophoresis of Al+ ions to the PVP GQD layer and the HfOx thin film. A local electric field increase occurred, resulting in the breakage of the conductive filament in the defect.

A chemical proteomic approach for in vivo evaluation of CCl4 effect on the acute serum proteins.

  • Uhm, Young-Ho;Kim, Young-Doo;Seo, Eun-Seok;Kang, Tae-Hyuk;Kang, Dong-Hoon;Soh, Yun-Jo;Kang, Chul-Hun
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.254.1-254.1
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    • 2003
  • CCl4 has been known as typical chemical which induce acute hepatitis accompanying increase in the levels of acute phase proteins in serum. In this study, after acute liver damage was induced by CCl4 in Sprague-Dawley rats, the levels of serum acute phase proteins were examined using 2-dimensional electrophoresis and lactic dehydrogenase (LDH), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in blood were also examined at 1, 2, 3 days after the induction to confirm acute hepatitis. (omitted)

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