| Project/Area Number |
15310141
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied genomics
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| Research Institution | Department of Physics, School of Science, Kitasato University |
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Principal Investigator |
KODERA Yoshio Kitasato University, Physics, Assistant Professor, 理学部, 講師 (60265733)
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| Co-Investigator(Kenkyū-buntansha) |
MAEDA Tadakazu Kitasato University, Physics, Professor, 理学部, 教授 (90265728)
OH-ISHI Masamichi Kitasato University, Physics, Assistant Professor, 理学部, 講師 (40233027)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥7,900,000 (Direct Cost: ¥7,900,000)
Fiscal Year 2004: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2003: ¥4,900,000 (Direct Cost: ¥4,900,000)
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| Keywords | proteomics / oxidative damage / protein / peptide / 蛍光検出 / ペプチドーム / ペプチド / プロテオーム解析 |
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| Research Abstract |
Reactive oxygen species (ROS) causes oxidative damages on nucleic acids, fatty acids, carbohydrates, and proteins in vivo. Side chains of lysine, arginine, proline and threonine residues are susceptible to ROS damages to be carbonylated. It has been demonstrated that protein carbonyls correlates well with aging and many diseases including diabetes, degenerative brain diseases (Alzheimer's disease and Parkinson's disease) and chronic lung disease. However, the relationships between the carbonylated proteins and those diseases have not yet been clarified.
Here, we established a method to mass-spectrometrically study the relationships between disease and protein carbonyls using an affinity tag called TOP (tags for oxidized proteins). TOP consists of three functional elements : an affinity tag, an isotope-coded linker, and hydrazide to react with protein carbonyls. The tag enables us to enrich carbonylated proteins from crude cell extracts and to subsequently identify carbonylated sites. Furthermore, TOP lets us quantitatively and directly analyze the protein carbonyls using mass spectroscopy.
We applied the TOP to studying states of artificially oxidized peptides by LC-MS and succeeded in purifying the carbonylated peptides from variously oxidized ones. It was found that carbonylation of lysine occurred after oxidation of methionine. Furthermore, this tag was applied to studying an artificially oxidized protein.
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