• 제목/요약/키워드: proteome analysis

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Proteome analysis of human stomach tissue: Separation of soluble Proteins by two-dimensional Polyacrylamide gel electrophoresis and identification by mass spectrometry

  • Ha, Geun-Hyoung;Lee, Seung-Uook;Kang, Deok-Gyeong;Ha, Na-Young;Kim, Soon-Hee;Kim, Ji-Na;Bae, Jong-Min;Kim, Jae-Won;Lee, Chang-Won
    • 한국생명과학회:학술대회논문집
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    • 한국생명과학회 2002년도 제38회 학술심포지움
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    • pp.20-47
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    • 2002
  • Two-dimensional gel electrophoresis (2-DE) maps for human stomach tissue proteins have been prepared by displaying the protein components of the tissue by 2-DE and identifying them using mass spectrometry. This will enable us to present an overview of the proteins expressed In human stomach tissues and lays the basis for subsequent comparative proteome analysis studies with gastric diseases such as gastric cancer. In this study, 2-DE maps of soluble fraction proteins were prepared on two gel images with partially overlapping pH ranges of 4-7 and 6-9. On the gels covering pH 4-7 and pH 6-9, about 900 and 600 protein spots were detected on silver staining, respectively. For protein identification, proteins spots on micropreparative gels stained by colloidal Coomassie Brilliant Blue G-250 were excised, digested in-gel with trypsln, and analyzed by peptide mass fingerprinting with delayed extraction-matrix assisted laser dosorption/ionization-mass spectrometry (DE-MALDI-MS). In all, 243 protein spots (168 spots in acidic map and 75 spots in basic map) corresponding to 136 different proteins were identified. Besides these principal maps, maps of lower resolution, i.e. overview maps (displayed on pH 3-10 gels) for total homogenate and soluble fraction, are also presented with some identifications mapped on them. Based on the 2-DE maps presented in this study, a 2-DE database for human stomach tissue proteome has been constructed and available at http://proteome.gsnu.ac.kr/DB/2DPAGE/Stomach/. The 2-DE maps and the database resulting from this study will serve important resources for subsequent proteomic studies for analyzing the normal protein variability in healthy tissues and specific protein variations in diseased tissues.

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Periodic Change in DO Concentration for Efficient Poly-${\beta}$-hydroxy-butyrate Production Using Temperature-inducible Recombinant Escherichia coli with Proteome Analysis

  • Abdul Rahman, Nor Aini;Shirai, Yoshihito;Shimizu, Kazuyuki;Hassan, Mohd Ali
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제7권5호
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    • pp.281-288
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    • 2002
  • Recombinant Escherichia coli strain harboring the ${\lambda}$pR-pL promotor and heterologus poly-${\beta}$-hydroxybutyrate (PHB) biosynthesis genes was used to investigate the effect of culture conditions on the efficient PHB production. The expression of phb genes was induced by a temperature upshift from $33^{\circ}C\;to\;38^{\circ}C$. The protein expression levels were measured by using two-dimensional electrophoresis, and the enzyme activities were also measured to understand the effect of culture temperature, carbon sources, and the dissolved oxygen (DO) concentration on the metabolic regulations. AcetylCoA is an important branch point for PHB production. The decrease in DO concentration lowers the citrate synthase activity, thus limit the flux toward the TCA cycle, and increase the flux for PHB production. Since NADPH is required for PHB production, the PHB production does not continue leading the overproduction of acetate and lac-tate. Based on these observations, a new operation was considered where DO concentration was changed periodically, and it was verified its usefulness for the efficient PHB production by experiments.

Proteome analysis of chloroplast proteins in stage albinism line of winter wheat (triticum aestivum) FA85

  • Hou, Dian-Yun;Xu, Hong;Du, Guang-Yuan;Lin, Jun-Tang;Duan, Min;Guo, Ai-Guang
    • BMB Reports
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    • 제42권7호
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    • pp.450-455
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    • 2009
  • The "stage albinism line of winter wheat" FA85 was a specific natural mutant strain on leaf color. This physiological mutation was controlled by cytogene. In order to reveal the genetic and biochemical mechanism of albinism, 2-DE was used to investigate the difference of chloroplast protein expression pattern between FA85 and its parent wheat Aibian 1. From the results of 2-DE gels analysis, approximately 683 spots were detected on each gel, and 57 spots were expressed differently at least two-fold. Using MALDI-TOF/TOF MS, 14 of 57 spots were identified, which could be categorized into four classes: carbon metabolism, energy metabolism, defense/stress response and signal transduction. Compared with the parent wheat, the expression of ATPase-$\gamma$ and GP1-$\alpha$ was up-regulated in FA85, and of other proteins was down-regulated. Together, we concluded that the expression of chloroplast proteins had changed obviously in FA85, which might be related to the leaf color mutant.