• Title/Summary/Keyword: 2D electrophoresis

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Clustering of 2D-Gel images (2H-Gel 이미지의 정렬 및 클러스터링)

  • Hur Won
    • KSBB Journal
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    • v.20 no.2 s.91
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    • pp.71-75
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    • 2005
  • Alignment of 2D-gel images of biological samples can visualize the difference of expression profiles and also inform us candidates of protein spots to be further analyzed. However, comparison of two proteome images between the case and control does not always successfully identify differentially expressed proteins because of sample-to-sample variation, poor reproducibility of 2D-gel electrophoresis and inconsistent electrophoresis conditions. Multiple alignment of 2D-gel image must be preceded before visualizing the difference of expression profiles or clustering proteome images. Thus, a software for the alignment of multiple 2D-Gel images and their clustering was developed by applying various algorithms and statistical methods. Microsoft Visual C++ was used to implement the algorithms in this work. Multiresoultion-multilevel algorithm was found out to be suitable for fast alignment and for largely distorted images. Clustering of 10 different proteome images of Fetal Alcohol Syndrome, was carried out by implementing a k-means algorithm and it gave a phylogenetic tree of proteomic distance map of the samples. However, the phylogenetic tree does not discriminate the case and control. The whole image clustering shows that the proteomic distance is more dependent to age and sex.

Purification and Characterization of D-Xylanase II from Penicillium verruculosum (Penicillium verruculosum으로부터 D-xylanase II의 정제 및 특성)

  • 조남철;강영태;이태훈;정기철;김강화
    • Microbiology and Biotechnology Letters
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    • v.21 no.6
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    • pp.588-593
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    • 1993
  • Xylanase(1, 4-beta-D-xylan xylanohydrolase` EC 3.2.1.8) II was purified from Penicillium verruculosum by using the techniques of two anion exchange chromatographies, and gel filtration. The molecular weight of this enzyme was about 22, 000 as determined by SDS-electrophoresis. The enzyme showed hydropytic activity toward xylan but did not catalyze hydrolysis of Rho-nitrophenyl-beta-D-xylopyranoside, Rho-nitrophenyl-beta-D-glucopyranoside, Rho-nitrophenyl-beta-D-cellobiopyranoside, and celluloses such as Avicel, cotton, filter paper, carboxymethylcellulose.

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Electrophoretic Behaviors of α-Lactalbumin and β-Lactoglobulin Mixtures Caused by Heat Treatment

  • Lee, You-Ra;Hong, Youn-Ho
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.7
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    • pp.1041-1045
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    • 2003
  • In order to study the reaction behaviors of bovine $\alpha$-lactalbumin ($\alpha$-La), $\beta$-lactoglobulin ($\beta$-Lg), and their mixtures during heat treatment, samples were analyzed using native-polyacrylamide gel electrophoresis (Native-PAGE), sodium dodecylsulfate (SDS)-PAGE, and two-dimensional (2-D)-PAGE. The electrophoresis demonstrated that the loss of native-$\alpha$-La increased as temperature increased, and that the loss of apo-$\alpha$-La was slightly higher than that of holo-$\alpha$-La. The tests also showed that during heat treatment, a mixture of $\alpha$-La and $\beta$-Lg was less stable than $\alpha$-La alone. As such, it was assumed that $\beta$-Lg induced holo-$\alpha$-La to be less stable than apo-$\alpha$-La during heat treatment. The reaction behavior of $\alpha$-La (holo-, apo-form) during heat treatment showed similar patterns in the 2-D-PAGE electropherogram, but the mixture of $\alpha$-La and $\beta$-Lg created new bands. In particular, the results showed a greater loss of native $\alpha$-La in the holo-$\alpha$-La and $\beta$-Lg mixture than in the apo-$\alpha$-La and $\beta$-Lg mixture. Thus, it can be concluded that the holo-$\alpha$-La and $\beta$-Lg mixture was more intensively affected by heat treatment than other samples, and that free sulphydryl groups took part in the heat-induced denaturation.

Protein Expression Analysis of Halobacillus dabanensis $D-8^T$ Subjected to Salt Shock

  • Feng De Qin;Zhang Bo;Lu Wei Dong;Yang Su Sheng
    • Journal of Microbiology
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    • v.44 no.4
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    • pp.369-374
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    • 2006
  • To investigate the mechanism of salt tolerance of gram-positive moderately halophilic bacteria, two-dimensional gel electrophoresis (2-D PAGE) was employed to achieve high resolution maps of proteins of Halobacillus dabanensis $D-8^T$. Approximately 700 spots of proteins were identified from these 2-D PAGE maps. The majority of these proteins had molecular weights between 17.5 and 66 kDa, and most of them were distributed between the isoelectric points (pI) 4.0 and 5.9. Some protein spots were distributed in the more acidic region of the 2-D gel (pI <4.0). This pattern indicated that a number of proteins in the strain $D-8^T$ are acidic. To understand the adaptation mechanisms of moderately halophilic bacteria in response to sudden environmental changes, differential protein profiles of this strain were investigated by 2-D PAGE and $Imagemaster^{TM}$ 2D Platinum software after the cells were subjected to salt shock of 1 to 25% salinity for 5 and 50 min. Analysis showed 59 proteins with an altered level of expression as the result of the exposure to salt shock. Eighteen proteins had increased expression, S proteins were induced, and the expression of 33 proteins was down-regulated. Eight of the up-regulated proteins were identified using MALDI-TOF/MS and MASCOT, and were similar to proteins involved in signal transduction, proteins participating in energy metabolism pathways and proteins involved in stress.

Proteome Analysis of Responses to Ascochlorin in LPS-induced Mouse Macrophage RAW264.7 Cells by 2-D Gel Electrophoresis and MALDI-TOF MS. (LPS로 자극된 macrophage RAW264.7 세포에서 ascochlorin에 대한 단백질체 분석)

  • Chang, Young-Chae
    • Journal of Life Science
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    • v.18 no.6
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    • pp.814-825
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    • 2008
  • Ascochlorin (ASC) is prenyl-phenol compound that was isolated from the fungus Ascochyta viciae. ASC reduces serum cholesterol and triglyceride levels, and suppresses hypertension, tumor development, ameliorates type I and II diabetes. Here, to better understand the mechanisms by which ASC regulates physiological or pathological events and induces responses in the pharmacological treatment of inflammation, we performed differential analysis of the proteome of the mouse macrophage RAW264.7 cells in response to ASC. In this study, we used a proteomic analysis of LPS-induced RAW264.7 cells treated by ASC, to identify proteins potentially involved in inflammatory processes. The RAW264.7 cell proteomes with and without treatment with ASC were compared using two-dimensional electrophoresis (2-D SDS-PAGE), matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF-MS) and bioinformatics. The largest differences in expression were observed for the calreticulin (4-fold decrease), ${\beta}-actin$ (4-fold decrease) and vimentin (1.5-fold decrease). In addition, rabaptin was increased 3-fold in RAW264.7 cells treated with ASC. The expression of some selected proteins was confirmed by RT-PCR analysis.

A Proteomic Approach for Quantitative Analysis of Calcitonin Gene-related Peptides in the Cerebrospinal Fluid Obtained from a Rat Model of Chronic Neuropathic Pain (만성 신경병성 통증이 유발된 쥐의 뇌척수액에서 단백체학을 이용한 Calcitonin Gene-related Peptides의 정량분석)

  • Kim, Dong Hee;Hong, Sung Ho
    • The Korean Journal of Pain
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    • v.21 no.2
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    • pp.112-118
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    • 2008
  • Background: This study was conducted to quantitatively analyze proteins associated with the calcitonin gene-related peptide (CGRP) in cerebrospinal fluid (CSF) that was obtained from a rat model of chronic neuropathic pain following administration of intrathecal $CGRP_{8-37}$. Methods: Male Sprague-Dawley rats (100-150 g, 5-6 wks) were divided into two groups, sham controls and neuropathic pain models. At the time of operation for neuropathic pain model, an intrathecal catheter was threaded through the intrathecal space. At 1 or 2 wks after the operation (maximum pain state), a test dose of 1, 5, 10, or 50 nM of $CGRP_{8-37}$ was injected into the intrathecal catheter and the CSF was then aspirated. Conventional proteomics to evaluate the CSF were then performed using high resolution 2-D, gel electrophoresis followed by computational image analysis and protein identification by mass spectrometry. Results: Treatment with $CGRP_{8-37}$ effectively alleviated mechanical allodynia in a dose dependent manner. The most effective response was obtained when a dose of 50 nM was administered, but significant differences were obtained following administration of only 5 nM $CGRP_{8-37}$. Furthermore, the results of the proteomic analysis were consistent with the experimental results. Specially we detected 30 differentially expressed spots in 7 images when 2-D gel electrophoresis was conducted. The intensity of 6 of these spots (spot number: 20 and 26-30) was found decrease the $CGRP_{8-37}$ dose increased; therefore, these spots were evaluated by mass spectrometry. This analysis identified 2 different proteins, CGRP (spot numbers: 26-30) and neurotensin-related peptide (spot number: 20). Conclusions: The results of this study suggest that CGRP plays a role in chronic central neuropathic pain and is a major target of chronic neuropathic pain management.

Identification of Rhizobium fredii and Bradyrhizobium japonicum by Polyacrylamide Gel Electrophoresis (전기영동법(電氣泳動法)에 의(依)한 대두(大豆) 근류균(根瘤菌) Rhizobium fredii와 Bradyrhizobium japonicum의 분류(分類) 및 동정(同定))

  • Yun, Han-Dae;Cho, Moo-Je;Lee, Ke-Ho
    • Applied Biological Chemistry
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    • v.30 no.2
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    • pp.163-168
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    • 1987
  • A method, based upon the separation of cellular proteins by one-and two-dimensional electrophoresis was used for distinguishing butween Bradyrhizobium japonicum strains and Rhizobium fredii strains. Significant differences in protein pattern of one-dimensional SDS-PAGE vs-ere observed between Rhizobium fredii strains and Bradyrhizobium japonicum strains. The differences in six distinct main lands were observed among total 52 kinds of protein bands. Furthermore, the distribution of proteins in two groups by two-dimensional polyacrylamide gel electrophoresis was very different. The majority of visible proteins of Rhizobium fredii were acidic, whereas those of Bradyrhizobium japonicum were basic. In addition, amino acid composition was analyzed to detect the differences between two groups. No significant differences in amino acid composition were observed between Bradyrhizobium japonicum strains and Rhizobium fredii strains. The results indicate that one-and two-dimensional polyacrylamide gel electrophoresis were useful for identifying rhizobia isolates. One-dimensional SDS-PAGE of rhizobia proteins provided a rapid method for screening a large number of isolates, whereas two-dimensional electrophoresis was more of resolution and easiness for analyzing protein spots.

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Impedance Matching of Electrically Small Antenna with Ni-Zn Ferrite Film

  • Lee, Jaejin;Hong, Yang-Ki;Lee, Woncheol;Park, Jihoon
    • Journal of Magnetics
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    • v.18 no.4
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    • pp.428-431
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    • 2013
  • We demonstrate that a partial loading of $Ni_{0.5}Zn_{0.5}Fe_2O_4$ (Ni-Zn ferrite) film remarkably improves impedance matching of electrically small $Ba_3Co_2Fe_{24}O_{41}$ ($Co_2Z$) hexaferrite antenna. A 3 ${\mu}m$ thick Ni-Zn ferrite film was deposited on a silicon wafer by the electrophoresis deposition process and post-annealed at $400^{\circ}C$. The fabricated Ni-Zn ferrite film has saturation magnetization of $268emu/cm^3$ and coercivity of 89 Oe. A partial loading of the Ni-Zn ferrite film on the $Co_2Z$ hexaferrite helical antenna increases antenna return loss to 24.7 dB from 9.0 dB of the $Co_2Z$ antenna. Experimental results show that impedance matching and maximum input power transmission to the antenna without additional matching elements can be realized, while keeping almost the same size as the $Co_2Z$ antenna size.