| Project/Area Number |
12204011
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| Research Category |
Grant-in-Aid for Scientific Research on Priority Areas
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| Allocation Type | Single-year Grants |
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| Review Section |
Biological Sciences
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| Research Institution | Kitasato University |
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Principal Investigator |
MAEDA Tadakazu Kitasato University, School of Science, Professor, 理学部, 教授 (90265728)
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| Co-Investigator(Kenkyū-buntansha) |
BABA Shiro Kitasato University, School of Science, Professor, 医学部, 教授 (00051889)
FUJITA Yoshikuni Kitasato University, School of Science, Professor, 医学部, 助教授 (50133286)
FURUDATE Sen-Ichi Kitasato University, School of Science, Professor, 医学部, 助教授 (80095512)
MATSUZAKI Takao Mitsubishi Chemical Corporation, Science and Technology Research Center, Fellow, 科学技術センター, フェロー
OMORI Akira Mitsubishi Kagaku Institute of Life Sciences, Laboratory Chief, 蛋白質構造解析室, 室長
大石 正道 北里大学, 理学部, 講師 (40233027)
亀谷 徹 北里大学, 医学部, 教授 (50101035)
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| Project Period (FY) |
2000 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥76,800,000 (Direct Cost: ¥76,800,000)
Fiscal Year 2004: ¥17,600,000 (Direct Cost: ¥17,600,000)
Fiscal Year 2003: ¥17,600,000 (Direct Cost: ¥17,600,000)
Fiscal Year 2002: ¥17,600,000 (Direct Cost: ¥17,600,000)
Fiscal Year 2001: ¥24,000,000 (Direct Cost: ¥24,000,000)
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| Keywords | Diabetes model animal / Protein expression profiling / Protein function profiling / Reactive oxygen species / Oxidatively damaged proteins / Tags for oxidized proteins / Disease peptidome analysis / Monoclonal antibodies for diagnosis / 発現プロテオミクス / 機能プロテオミクス / 抗体基盤のプロテオミクス / 疾患ペプチドミクス / 酸化ストレス / 酸化傷害タンパク質の蓄積 / ユビキチン化 / プロテオーム解 / 糖尿病 / ヒト組織バンク / タンパク質機能状態 / プロテオーム解析 / 糖尿病特異的プロテオーム / ob / obマウス / db / dbマウス / OLETFラット / ヒト標品のプロテオーム解析 / ヒト・ティッシュ・バンク / 電気泳動画像データベース |
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| Research Abstract |
(1) Setting up a proteomics research environment; We use a two-dimensional electrophoresis (2-DE) method far separating proteins and LC/MS for identifying the proteins. We can routinely identify 90 to 95% of proteins separated by the 2-DE and visualized by Commassie staining. Speed of identification is 80 proteins per day.
(2) Protein expression profiling of various organs of diabetes model rats: Protein expression profiling was carried out for the diabetes model OLETF and control LETO rats. The OLETF rats on average develop diabetes at 20 weeks of age, but meaningful differences were not found between expression proteomes of the rats with and without the disease.
(3) Proteome-wide detection of proteins oxidatively damaged by the reactive oxygen species (ROS): Anew method was developed for detecting the ROS damaged proteins. Application of the method revealed that ROS-damaged proteins increased in the left ventricle of the OLETF rats as the diabetes proceeded.
(4) Tags for Oxidized Proteins (TOP): New tags for the oxidatively damaged proteins were developed suitable for concentrating minute amount of proteins and for analyzing the proteins by HPLC and/or MS.
(5) Disease peptidome based on a new extraction method of peptides from tissues and blood sera: A highly efficient method was developed for extracting peptides in tissues and blood sera. Application of the method let us find cancer marker peptides in blood sera of cancer patients.
(6) Obtaining monoclonal antibodies useful for disease diagnosis: Hirohashi, et al., obtained a monoclonal antibody by means of a unique immunizing procedure (Gann, 75, 485-488, 1984). We extended their procedure to obtain monoclonal antibodies useful for disease diagnosis. By using the procedure we have successfully identified several antibodies and antigens useful for cancer diagnosis.
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