生物物理化学
Online ISSN : 1349-9785
Print ISSN : 0031-9082
ISSN-L : 0031-9082
47 巻, 1-2 号
選択された号の論文の7件中1~7を表示しています
  • 戸田 年総
    2003 年 47 巻 1-2 号 p. 1-6
    発行日: 2003/06/15
    公開日: 2009/03/31
    ジャーナル フリー
    The comprehensive analysis of post-translational modification of protein is thought to be the most important object of biomedical proteomics in the post-genome era. Especially, protein phosphorylation is quite important process in the physiological regulation of metabolism, and the irregular phosphorylation might be resulted in various diseases including malignant tumors. However, the concentration of those regulatory phosphoprotein level is generally too low to be detected directly by the mass spectrometry in proteomics. Therefore, preliminary concentration of phosphopeptides by the method of immobilized iron affinity chromatography (IMAC) is necessary to achieve the sensitive detection. In this paper, the present status and the prospect of the method of IMAC are discussed.
  • 荒木 令江, 川野 克己, 戸田 年総, 荒木 朋洋, 次田 晧, 吉良 潤一, 佐谷 秀行
    2003 年 47 巻 1-2 号 p. 7-16
    発行日: 2003/06/15
    公開日: 2009/03/31
    ジャーナル フリー
    More than three hundreds of genes related to brain diseases have been identified in human genome analysis. However, there are few information on the gene products (proteins) that give us the most important idea on the biological mechanism of the disease formation. Brain proteomics have been concentrated on the disease related to the brain tumor and neuronal apoptosis that involve many regulator proteins for cellular processes, such as cell cycle, DNA repair, apoptosis, etc. Many factors contribute to control their activations, and their downstream responses also depend on the cellular environment or other modifying factors in the cells of the central nervous system (CNS). To compile the information of the brain protein structures and functions related to the pathogenesis, and to make use of the information to the clinical application, proteomic study for the brain tissue/cells must be crucial. We have been constructing proteome maps of mouse and human brain by two-dimensional electrophoresis (2-D) and listing the information of protein expression profiles in the data bank. In this review, our proteomic approaches on the specific brain tissue and cellular proteins that relate to the neuronal apoptosis and brain tumor were introduced with two functional proteomic strategies; 1) Differential proteomic display (comparative 2-D) of brain tissue proteins, 2) Affinity cellular proteomic mapping; detection of specific binding proteins that associate with the target gene products using micro affinity columns. Our final goal on this study is to contribute to the advancement of medical science as well as brain science by constructing the proteomic database covered the gamut of protein information of the CNS disease related concerns.
  • 友杉 直久
    2003 年 47 巻 1-2 号 p. 17-22
    発行日: 2003/06/15
    公開日: 2009/03/31
    ジャーナル フリー
    Needle biopsy is the standard test for the diagnosis of renal diseases. Biopsy-associated complications could not be eliminated in spite of recent refinement. The deveopment of noninvasive diagnostic test that provides insights into the mechanisms of renal diseases would be expected. Recently the advent of SELDI-TOF-MS (surface-enhanced laser desorption/ionization time-of-flight mass spectrometry) has extended the application of mass spectrometry to the study of proteins from complex biological systems. We applied the new protein-chip technology based on SELDI in a discovery of renal disease biomarkers. Proteomic patterns in serum by means of protein-chip were exemplified by elucidating a biomarker candidate for acute renal allograft rejection. In discovery phase protein profiles for control and rejection were compared in protein expression. The process of characterization and validation for the biomarker could be monitored by MS detection. SELDI protein-chip technology will be applied more frequently to a number of medical and basic research problems because of high resolution, high reproducibility, ease of use, and fentomole sensitivity.
  • 真鍋 敬
    2003 年 47 巻 1-2 号 p. 23-26
    発行日: 2003/06/15
    公開日: 2009/03/31
    ジャーナル フリー
    Two-dimensional polyacrylamide gel electrophoresis (2-D PAGE), followed by protein extraction and characterization with chemical sequencing or mass spectrometry (MS), is the most commonly used method to analyze complex protein systems such as cells and organella. Many reports appeared on the improvements of these processes of information-collection, to establish a system with higher performance. However, it is claimed that 2-D PAGE-based methods are slow and labor-intensive and also need subsequent efforts for one-by-one identification of proteins. Combined methods of Fourier transform ion cynclotron resonance (FTICR) mass spectrometry with preceding separation techniques such as capillary isoelectric focusing or capillary liquid chromatography was introduced to be promising for the analysis of the protein systems. Also, the works to overcome the disadvantages of 2-D PAGE-based methods, combining the results obtained by non-denaturing 2-D PAGE and by denaturing 2-D PAGE to integrate protein information, were introduced.
  • 勝田 和大, 野村 英子, 稲垣 直之
    2003 年 47 巻 1-2 号 p. 27-31
    発行日: 2003/06/15
    公開日: 2009/03/31
    ジャーナル フリー
    A fundamental concern about proteomics is improving the coverage of detection of a cellular proteome using our technology. In the case of two-dimensional gel electrophoresis (2-DE), a core technology of proteomics, enlargement of the gel size appears a straightforward and effective strategy for increasing the number of detected proteins. We utilized fourteen large 2-DE gels (twelve 24cm×7cm gels) which were assembled into a 93cm×103cm cybergel. Our data suggested that the cyber gel can display more than 11, 000 protein spots expressed in a 1-105 dynamic range in cells.
  • Nazrul Islam, 平野 久
    2003 年 47 巻 1-2 号 p. 33-42
    発行日: 2003/06/15
    公開日: 2009/03/31
    ジャーナル フリー
    Current knowledge of protein-protein interactions in proteome is very limitted due to the lack of high throughput and efficient techniques at the protein level. In this perspective, we briefly discuss the techniques currently available to determine protein-protein interactions and then focus on a novel technique developed by manipulating genome composition in common wheat. Genome manipulation has been carried out either by deleting chromosomes-an arm (ditelocentric lines), a part of chromosome (fine deletion), one of the three genomes, A, B, D-or by introducing a pair of chromosomes (disomic addition line) in the genome composition of common wheat. The network of protein-protein interactions in the wheat seed proteome was determined by quantitative analysis of the expressed proteins. Protein expression in the proteome was found to be controlled by a network. Deletion of chromosome(s), either an arm, a part of an arm or one of the three genomes, has resulted a serious imbalance in the network of protein expression. Addition of extra chromosomes, hence proteins, from the alien species did not affect much to the net work of protein expression. This result indicates that the expression of proteins in proteome is not totally independent, rather there is a network of proteins which determines protein expression in the proteome, and the deletion of chromosome(s), hence proteins, from the network causes a serious irregularities in protein expression as compared to the addition.
  • Nazrul Islam, 高岡 素子, 佐々 英徳, 川崎 博史, 平野 久
    2003 年 47 巻 1-2 号 p. 43-49
    発行日: 2003/06/15
    公開日: 2009/03/31
    ジャーナル フリー
    Starch granule proteins (SGPs) were extracted from wheat seeds and characterized using proteomic approaches: two dimensional electrophoresis (2-DE), 40cm long gradient sodium dodecyl sulfate (SDS) gel electrophoresis and electrospray ionization mass spectrometry. The majority of the low molecular weight (LMW) SGPs, especially in the range of 14-30kDa, was located on the surface of starch granule, and only a small number of LMW SGPs of molecular weight range from 30 to 55kDa was found within starch granules. The surface SGPs were mostly expressed in acidic, whereas the integral SGPs were expressed basic regions of 2-DE. Gene location of one surface SGP of approximately 14kDa was assigned on short arm of chromosome 4B. One of the LMW SGPs was identified by LC-ESI MS/MS endochitinase (30kDa). Intensity of this proteins was found to decrease during seed germination and seed maturation.
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