2004 Fiscal Year Final Research Report Summary
Tags for oxidized proteins (TOP) useful for assessing the effect of reactive oxygen species on disease proteomics
| 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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| Keywords | proteomics / oxidative damage / protein / peptide |
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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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[Journal Article] Curative effect of thyroxine treatment on pancreatic proteomes of the rdw rat having hereditary hypothyroidism.2005
Author(s)
Satoh, M., Sato-Haruta, E., Omori, A., Oh-Ishi, M., Kodera, Y., Furudate, S-I., Maeda, T.
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Journal Title
Proteomics 5
Pages: 1113-1124
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] High molecular mass proteome of androgen independent prostate cancer.2005
Author(s)
Kuruma, H., Egawa, S., Oh-Ishi, M., Kodera, Y., Satoh, M., Chen, W., Satoh, T., Baba, S.
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Journal Title
Proteomics 5
Pages: 1097-1112
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] A simple method for peptide purification as a basis for peptidome analysis.2005
Author(s)
Fukutomi, T., Kodera, Y., Kogo, T., Furudate, S., Omori, A., Maeda, T.
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Journal Title
J.Electrophoresis 49
Pages: 15-21
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Solution structure and functional characterization of SGTx1, a modifier of Kv2.1 channel gating.2004
Author(s)
Lee CW, Kim S, Roh SH, Endoh H, Kodera Y, Maeda T, Kohno T, Wang JM, Swartz KJ, Kim JI
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Journal Title
Biochemistry. 43
Pages: 890-897
Description
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[Journal Article] Identification of altered protein expression and post-translational modifications in primary colorectal cancer by using agarose two-dimensional gel electrophoresis.2004
Author(s)
Tomonaga, T., Matsushita, K., Yamaguchi, S., Oh-Ishi, M., Kodera, Y., Maeda, T., Shimada, H., Ochiai, T., Nomura, F.
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Journal Title
Clin.Cancer Res. 10
Pages: 2007-2014
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Solution structure and functional characterization of SGTx1, a modifier of Kv2.1 channel gating.2004
Author(s)
Lee, C.W., Kim, S., Roh, S.H., Endoh, H., Kodera, Y., Maeda, T., Kohno, T., Wang, J.M., Swartz, K.J., Kim, J.I.
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Journal Title
Biochemistry 43
Pages: 890-897
Description
「研究成果報告書概要(欧文)」より
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