• Title/Summary/Keyword: two-dimensional gel

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Proteomic analysis of differentially expressed skin proteins in iRhom2Uncv mice

  • Liu, Bing;Xu, Yuan;Li, Wen-Long;Zeng, Lin
    • BMB Reports
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    • v.48 no.1
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    • pp.19-24
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    • 2015
  • A mouse homozygous for the spontaneous mutation uncovered (Uncv) has a hairless phenotype. A 309-bp non-frameshift deletion mutation in the N-terminal cytoplasmic domain of iRhom2 was identified in Uncv mice ($iRhom2^{Uncv}$) using target region sequencing. The detailed molecular basis for how the iRhom2 mutation causes the hairless phenotype observed in the homozygous $iRhom2^{Uncv}$ mouse remains unknown. To identify differentially expressed proteins in the skin of wild-type and homozygous $iRhom2^{Uncv}$ littermates at postnatal day 5, proteomic approaches, including two-dimensional gel electrophoresis and mass spectrometry were used. Twelve proteins were differentially expressed in the skin in a comparison between wild-type and homozygous $iRhom2^{Uncv}$ mice. A selection of the proteomic results were tested and verified using qRT-PCR, western blot and immunohistochemistry. These data indicate that differentially expressed proteins, especially KRT73, MEMO1 and Coro-1, might participate in the mechanism by which iRhom2 regulates the development of murine skin.

Plasma Protein Adsorption to Anion Substituted Poly(vinyl alcohol) Membranes

  • Ryu, Kyu-Eun;Hyangshuk Rhim;Park, Chong-Won;Chun, Heung-Jae;Hong, Seung-Hwa;Kim, Jae-Jin;Lee, Young-Moo
    • Macromolecular Research
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    • v.11 no.6
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    • pp.451-457
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    • 2003
  • Anion-substituted poly(vinyl alcohol) (PVA) membranes, carboxymethylated PVA (C-PVA), and sulfonated PVA (S-PVA) were prepared and the effects of these substitutions on the plasma protein adsorption were studied by one- and two-dimensional gel electrophoresis and immunoblotting. When Cuprophane was used as a negative control, the amount of total proteins bound to samples decreased in the order Cuprophane > PVA > C-PVA > S-PVA, which we attribute to the effects of the surface characteristics of the samples, such as their surface tensions and electrostatic properties, on the adsorption of proteins to the surfaces of the materials. The results revealed that albumin was the most abundant protein in all the samples. The proportion of adsorbed fibrinogen to S-PVA exceeded those of PVA and C-PVA, whereas S-PVA exhibited the lowest IgG adsorption affinity among the samples we studied.

Proteomic Analysis of Pancreata from Mini-Pigs Treated with Streptozotocin as Type I Diabetes Models

  • Lee, Phil-Young;Park, Sung-Goo;Kim, Eun-Young;Lee, Myung-Sup;Chung, Sang-J.;Lee, Sang-Chul;Yu, Dae-Yeul;Bae, Kwang-Hee
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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    • pp.817-820
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    • 2010
  • Type 1 diabetes mellitus (T1DM) is an autoimmune disease characterized by extreme insulin deficiency due to an overall reduction in the mass of functional pancreatic ${\beta}$-cells. Several animal models have been used to study T1DM. Amongst these, the mini-pig seems to be the most ideal model for diabetes research, owing to similarities with humans in anatomy and physiology. The purpose of this study was to analyze differentially expressed pancreatic proteins in a streptozotocin (STZ)-induced mini-pig T1DM model. Pancreas proteins from mini-pigs treated with STZ were separated by two-dimensional gel electrophoresis, and 11 protein spots were found to be altered significantly when compared with control mini-pigs. The data in this study utilizing proteomic analysis provide a valuable resource for the further understanding of the T1DM pathomechanism.

Changes in the RNA and Protein Synthesis at the Pre- and Post fertilization Stages of a Sea Urchin, Hemicentrotus pulcherrimus (말똥성게 (Hemicentrotus polcherrimus)의 수정전과 초기 발생동안 RNA 및 단백질합성의 변화)

  • Jang, Jeong-Won;Lee, Yang-Rim
    • The Korean Journal of Zoology
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    • v.28 no.2
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    • pp.71-84
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    • 1985
  • Syntheses of RNA and protein were studied to examine changes in activating stored mRNAs during the early development of a sea urchin, Hemicentrotus pulcherimus. The rates of RNA and protein syntheses are very low in the unferilized eggs but the protein synthesis is activated upon the fertilization, while RNA synthesis remains still inactive at the same stage. These rates increase drastically at the blastula and gastrula stages, although the increases are not exactly in parallel. The protein synthesis was found to be also changing in quality during the early development from the studies by the two-dimensional gel electrophoresis.

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The Effect of NaCI Treatment on the Freezing Tolerance and Protein Patterns of Carrot Callus Suspension Culture

  • Moon, Soon-Ok;Park, Sook-Hee;Cho, Bong-Heuy
    • BMB Reports
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    • v.30 no.1
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    • pp.21-25
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    • 1997
  • The growth. freezing resistance and electrophoretic protein patterns of carrot callus cultures were investigated following treatment with NaCl for various' intervals at 20$^{\circ}C$. Following 7 day exposure to 250 mM NaCl. freezing tolerance increased, which was measured by 2.3.5-triphenyl tetrazolium chloride (TTC) assay and fresh weight was reduced compared to control cells. Changes of electrophoretic patterns of total and boiling stable proteins were investigated using one or two dimensional gel system. Several proteins with molecular weight of 43 and 21 kDa increased by NaCl treatment. The most prominent change was detected in 21 kDa protein. The steady state level of this protein increased in NaCl treated cells, but decreased in control cells. Twenty one kDa protein was detected only in the NaCl treated cell when boiling stable protein was analyzed. The isoelectric point of 21 kDa protein was identified as 5.7. The timing of increase of 21 kDa protein was correlated to freezing resistance which implied the role of this protein in the induction of freezing resistance of the cell.

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Proteome Analysis of Waito-c Rice Seedlings Treated with Culture Fluid of Gibberellin-producing Fungus, Fusarium proliferatum KGL0401

  • Rim, Soon-Ok;Lee, Jin-Hyung;Hwang, Seon-Kap;Suh, Seok-Jong;Lee, Jin-Man;Rhee, In-Koo;Kim, Jong-Guk
    • Journal of Microbiology and Biotechnology
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    • v.16 no.12
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    • pp.1990-1994
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    • 2006
  • Fusarium proliferatum KGL0401 was previously isolated from Physalis alkekengi var. francheti plant roots and exhibited a high GA productivity. A gas chromatography-mass spectrometry (GC-MS) analysis of extracts of the culture fluid of F proliferatum KGL0401 also revealed the presence of $GA_1$, $GA_3$, $GA_4$, $GA_7$, $GA_{20}$, and $GA_{24}$. Therefore, the present study conducted a proteome analysis of waito-c rice treated with the culture fluid of the isolated F proliferatum KGL0401 to identify the protein expression triggered by the GA-containing culture fluid. The results revealed the overexpression of 180 protein spots in the sample treated with the culture fluid. Among them, 75 induced proteins were selected and analyzed by MALDI-TOF (matrix-assisted laser desorption-iorrization time-of-flight) mass spectrometry, followed by database searching, and 51 proteins were identified.

Interactome Analysis of Yeast Glutathione Peroxidase 3

  • Lee, Phil-Young;Bae, Kwang-Hee;Kho, Chang-Won;Kang, Sung-Hyun;Lee, Do-Hee;Cho, Sa-Yeon;Kang, Seong-Man;Lee, Sang-Chul;Park, Byoung-Chul;Park, Sung-Goo
    • Journal of Microbiology and Biotechnology
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    • v.18 no.8
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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.

Immunochemical Studies on Expression of Quinoproteins in Escherichia coli

  • Ryou, Chong-Suk;Kim, Jae-Beom;Kwon, Moo-Sik
    • Journal of Microbiology and Biotechnology
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    • v.10 no.1
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    • pp.95-98
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    • 2000
  • An immunochemical method has been develooped as the most sensitive tool for studying the expression of quinoproteins containing pyrroloquinoline qinone(PQQ) in E. coli. The PQQ was conjugated to bovine serum albumin (BSA), and the conjugant was purified by using a $KwikSep^{TM}$ dextran desalting column chromatography. The PQQ-BSA conjugant was immunized to rabbits, and the IgG fractions of the antisera were purified. The most sensitive antibody against PQQ-BSA conjugant recognized some nanogram quantity of the antigen on the blot, but had little cross reactivity with BSA. Using this batch of the antibody, all the immunochemical assays of quinoproteins in E. coli were preformed. Some six different PQQ-specfic spots were detected by Western blot analysis of the soluble proteins in E. coli were performed. Some six different PQQ-specific spots were detected by Western blot analysis of the soluble proteins in E. coli after two-dimensional gel electrophoresis. Their molecular weights on the blot were estimated to be about 100-, 90-, 72-, 58-, 52-, and 50kDa. Their pI values fell in the range from 4.8 to 5.5. These results stronly suggest that quinoproteins are present in E. coli, and that the protein moieties were covalently bound to PQQ.

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THE FINITE ELEMENTS ANALYSIS IN THE THREE DIMENSIONAL CELL CULTURE MODEL OF THE COLLAGEN MATRIX ACCORDING TO THE APPLICATION FORCE (교원섬유 기질을 이용한 3차원 세포 배양 모델에 기계적인 힘을 가하는 경우 기질을 통하여 세포에 가해지는 힘의 특성에 대한 비교 분석 연구)

  • Kim, Hyeon-Jong;Rhyu, In-Chul;Park, Jun-Woo;Kim, Seong-Gon
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.34 no.2
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    • pp.151-156
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    • 2008
  • Different kinds of forces can be applied to the biological tissue. The analysis of the applied force is highly important to explain the mechanism of cellular response. In this study, the applied force to the collagen gel was analyzed by the finite elements analysis. The model received two different kinds of static force (compression and tension). The force range was 50g to 400g. In results, von Mises stress was concentrated in the peripheral region in the compression model. It was concentrated in the central area in the tension model. However, the compressive force was high in the peripheral area of the compression model and the tensional force was also high in the same area of the tension model. In conclusion, the applied force could be different to the region and it should be considered in the experiment to analyze the effects of the mechanical force on the cells.

Tissue proteomics for cancer biomarker development - Laser microdissection and 2D-DIGE -

  • Kondo, Tadashi
    • BMB Reports
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    • v.41 no.9
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    • pp.626-634
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    • 2008
  • Novel cancer biomarkers are required to achieve early diagnosis and optimized therapy for individual patients. Cancer is a disease of the genome, and tumor tissues are a rich source of cancer biomarkers as they contain the functional translation of the genome, namely the proteome. Investigation of the tumor tissue proteome allows the identification of proteomic signatures corresponding to clinico-pathological parameters, and individual proteins in such signatures will be good biomarker candidates. Tumor tissues are also a rich source for plasma biomarkers, because proteins released from tumor tissues may be more cancer specific than those from non-tumor cells. Two-dimensional difference gel electrophoresis (2D-DIGE) with novel ultra high sensitive fluorescent dyes (CyDye DIGE Fluor satulation dye) enables the efficient protein expression profiling of laser-microdissected tissue samples. The combined use of laser microdissection allows accurate proteomic profiling of specific cells in tumor tissues. To develop clinical applications using the identified biomarkers, collaboration between research scientists, clinicians and diagnostic companies is essential, particularly in the early phases of the biomarker development projects. The proteomics modalities currently available have the potential to lead to the development of clinical applications, and channeling the wealth of produced information towards concrete and specific clinical purposes is urgent.