Tags for oxidized proteins (TOP) useful for assessing the effect of reactive oxygen species on disease proteomics

• Project/Area Number: 15310141
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied genomics
• Research Institution: Department of Physics, School of Science, Kitasato University
• Principal Investigator: KODERA Yoshio Kitasato University, Physics, Assistant Professor, 理学部, 講師 (60265733)
• 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)
• Project Period (FY): 2003 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥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)
• 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

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.

Research Products

• [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
  • Journal Title: J.Electrophoresis 49 Pages: 15-21
  • NAID: 10015570325
  • Description: 「研究成果報告書概要(和文)」より
• [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
  • Journal Title: Proteomics 5 Pages: 1097-1112
  • NAID: 130006995824
  • Description: 「研究成果報告書概要(和文)」より
• [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
  • Journal Title: Proteomics 5 Pages: 1113-1124
  • Description: 「研究成果報告書概要(和文)」より
• [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.
  • Journal Title: Proteomics 5 Pages: 1113-1124
  • Description: 「研究成果報告書概要(欧文)」より
• [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.
  • Journal Title: Proteomics 5 Pages: 1097-1112
  • NAID: 130006995824
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Proteome analysis of prostate cancer (2005)
  • Author(s): Kuruma, H., Egawa, S., Oh-Ishi, M., Kodera, Y., Maeda, T.
  • Journal Title: Prostate Cancer and Prostatic Disease 8 Pages: 14-21
  • Description: 「研究成果報告書概要(欧文)」より
• [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.
  • Journal Title: J.Electrophoresis 49 Pages: 15-21
  • NAID: 10015570325
  • Description: 「研究成果報告書概要(欧文)」より
• [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
  • Journal Title: Clin.Cancer Res. 10 Pages: 2007-2014
  • NAID: 130006995830
  • Description: 「研究成果報告書概要(和文)」より
• [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
  • Journal Title: Biochemistry. 43 Pages: 890-897
  • Description: 「研究成果報告書概要(和文)」より
• [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.
  • Journal Title: Clin.Cancer Res. 10 Pages: 2007-2014
  • NAID: 130006995830
  • Description: 「研究成果報告書概要(欧文)」より
• [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.
  • Journal Title: Biochemistry 43 Pages: 890-897
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Low pKa lysine residues at the active site of sarcosine oxidase from Colynebacterium sp.U-96 (2004)
  • Author(s): Mukouyama, E.B., Oguchi, M., Kodera, Y., Maeda, T., Suzuki, H.
  • Journal Title: Biochem.Biophys.Research Comm. 320 Pages: 846-851
  • Description: 「研究成果報告書概要(欧文)」より
• [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, Ochi
  • Journal Title: Clin.Cancer Res. 10 Pages: 2007-2014
  • NAID: 130006995830
• [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 TT
  • Journal Title: Biochemistry 43 Pages: 890-897
• [Journal Article] Low pKa lysine residues at the active site of sarcosine oxidase from Corynebacterium sp. U-96 (2004)
  • Author(s): Mukouyama E.B., Oguchi M., Kodera Y., Maeda T., Suzuki H
  • Journal Title: Biochem.Biophys.Research Comm. 320 Pages: 846-851
• [Journal Article] Proteomic method detects oxidatively induced protein carbonyls in muscles of a diabetes model Otsuka Long-Evans Tokushima Fatty (OLETF) rat. (2003)
  • Author(s): Oh-Ishi M, Ueno T, Maeda T
  • Journal Title: Free Radic.Biol.Med 34 Pages: 11-24
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Proteomic method detects oxidatively induced protein carbonyls in muscles of a diabetes model Otsuka Long-Evans Tokushima Fatty(OLETF) rat. (2003)
  • Author(s): Oh-Ishi, M., Ueno, T., Maeda, T.
  • Journal Title: Flee Radic.Biol.Med. 34 Pages: 11-22
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Proteome analysis of prostate cancer.
  • Author(s): Kuruma, H, Egawa, S, Oh-Ishi, M, Kodera, Y, Maeda, T.
  • Journal Title: (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Proteomic method detects oxidatively induced protein carbonyls in muscles of a diabetes model Otsuka Long-Evans Tokushima Fatty (OLETF) rat.
  • Author(s): Fukutomi T, Kodera Y, Kogo T, Furudate S.Omori A, Maeda T
  • Journal Title: Journal of Electrophoresis (in press)
• [Journal Article] High molecular mass proteome of andorogen-independent pros tate cancer
  • Author(s): Kuruma H, Egawa S, Oh-Ishi M, Kodera Y, Satoh M, Chen W, Okusa H, Matsumoto K, Maeda T, Baba S
  • Journal Title: Proteomics (in press)
• [Journal Article] Effect of thyroxine on abnormal proteomes of the hypothyroi rdw rat
  • Author(s): Satoh M, Haruta-Satoh E, Omori A, Oh-Ishi M, Kodera Y, Furudate S, Maeda T
  • Journal Title: Proteomics (in press)
• [Book] ゲノム医科学解析 ゲノミクス・プロテオミクスの新展開 (2005)
  • Author(s): 前田忠計, 大石正道, 小寺義男
  • Total Pages: 7
  • Description: 「研究成果報告書概要(和文)」より
• [Patent(Industrial Property Rights)] 標的物質濃縮用機能性タグ、及び該機能性タグの使用方法 (2004)
  • Inventor(s): 小寺 義男, 前田 忠計
  • Industrial Property Rights Holder: JST
  • Industrial Property Number: 2004-105405
  • Filing Date: 2004-03-31
  • Description: 「研究成果報告書概要(和文)」より
• [Patent(Industrial Property Rights)] 酸化傷害タンパク質解析用タグ、及び該タグを用いる酸化傷害タンパク質の検出方法 (2004)
  • Inventor(s): 小寺 義男, 前田 忠計
  • Industrial Property Rights Holder: JST
  • Industrial Property Number: 2004-133054
  • Filing Date: 2004-04-28
  • Description: 「研究成果報告書概要(和文)」より
• [Patent(Industrial Property Rights)] 酸化傷害タンパク質解析用タグ、及び該タグを用いる酸化傷害タンパク質の検出方法 (2004)
  • Inventor(s): 小寺義男, 前田忠計
  • Industrial Property Rights Holder: 独立行政法人科学技術振興機構
  • Industrial Property Number: 2004-133054
  • Filing Date: 2004-04-28
• [Publications] Lee CW, Kim S, Roh SH, Endoh H, Kodera Y, Maeda T, Kohno T, Wang JM, Swartz KJ, Kim JI.: "Solution structure and functional characterization of SGTx1,a modifier of Kv2.1 channel gating"Biochemistry. 43. 890-897 (2004)
• [Publications] Kuruma H, Egawa S, Oh-Ishi M, Kodera Y, Maeda T: "Proteome analysis of prostate cancer"Asian J.Androl. (In press). (2004)
• [Publications] 0h-Ishi M, Ueno T, Maeda T: "Proteomic method detects oxidatively induced protein carbonyls in muscles of a diabetes model Otsuka Long-Evans Tokushima Fatty (OLETF) rat"Free Radic.Biol.Med.. 34. 11-22 (2003)
• [Publications] 前田忠計, 大石正道, 上野剛, 小寺義男: "糖尿病ラット筋肉における酸化傷害タンパク質の検出"J.Mass Spectrom.. 51. 509-515 (2003)
• [Publications] 大石正道, 小寺義男, 前田忠計: "遺伝性疾患モデル動物を用いた疾患プロテオーム解析"J.Mass Spectrom.. 51. 516-523 (2003)
• [Publications] 前田忠計, 大石正道, 小寺義男: "ゲノミクス・プロテオミクスの新展開(監修:今中忠行)"(株)エヌ・ティー・エス. (2004)