Identification of human mitochondrial metabolome proteins

• Project/Area Number: 15390106
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Pathological medical chemistry
• Research Institution: KYUSHU UNIVERSITY
• Principal Investigator: KANG Dongchon Kyushu University, Graduate School of Medical Sciences, Associate Professor, 大学院・医学研究院, 助教授 (80214716)
• Project Period (FY): 2003 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥15,400,000 (Direct Cost: ¥15,400,000); Fiscal Year 2004: ¥5,900,000 (Direct Cost: ¥5,900,000); Fiscal Year 2003: ¥9,500,000 (Direct Cost: ¥9,500,000)
• Keywords: mitochondria / mitochondrial DNA / nucleoid / mass spectrometery / mitochondrial transcription factor A / TFAM / RNAi / ヌクレオイド / マススペクトラム / メタボリオーム

Research Abstract

(Aim) Mitochondrial DNA (mtDNA) encodes genes essential for the mitochondrial electron transfer system. Recently we proposed that mtDNA takes on a large protein-DNA complex called nucleoid and the nucleoid structure is critical for maintenance of mtDNA. Mitochondrial transcription factor A (TFAM) is a main component of the nucleoid. In this study, I aim to identify protein components of the nucleoid by LC/MS/MS.
(Methods) Isolated human mitochondria were treated with a c ross-linker. After solubilization, the nucleoid was immunoprecipitated with anti-TEAM antibody. The precipitated proteins were separated on one dimensional SDS-PAGE, extracted, tripsinized, and and analyzed by LC/MS/MS.
(Results)
1)Potential DNA- or RNA binding proteins
2)Heat shock proteins
3)Ribosomal proteins
4)TCA cycle related proteins
5)Enzymes for β oxidation of fatty acids
All proteins identified were classified into only 5 groups as above. These results suggest that the mitochondrial nucleoid provides a scaffold for efficient metabolism of translation, TCA cycles, and β oxidation of fatty acids.

Research Products

• [Journal Article] Alterations of Mitochondrial DNA in common Diseases and Disease States (2005)
  • Author(s): Kang, D.
  • Journal Title: Curr.Med.Chem. 12 Pages: 429-441
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Mitochondrial transcription factor A ub the maintaenance of mtiochondrial DNA : Overview of its multiple role (2005)
  • Author(s): Kang, D.
  • Journal Title: Ann N Y Acad Sci 1042 Pages: 1-8
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Alterations of Mitochondrial DNA in Common Diseases and Disease States : Aging, Neurodegeneration, Heart Failure, Diabetes and Cancer. (2005)
  • Author(s): Kang, D., Hamasaki, N.
  • Journal Title: Curr.Med.Chem. 12 Pages: 429-441
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Mitochondrial transcription factor A in maintenance of mitochondrial DNA. (2005)
  • Author(s): Kang, D., Hamasaki, N.
  • Journal Title: Ann N Y Acad Sci 1042 Pages: 1-8
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Alterations of Mitochondrial DNA in Common Diseases and Disease States (2005)
  • Author(s): Kang, D.
  • Journal Title: Curr.Med.Chem. 12 Pages: 429-411
• [Journal Article] Mitochondrial transcription factor A ub the maintenance of mitochondrial DNA : Overview of its multiple role (2005)
  • Author(s): Kang, D.
  • Journal Title: Ann N Y Acad Sci 1042 Pages: 1-8
• [Journal Article] Architectural role of TFAM in maintenance of human mitochondrial DNA (2004)
  • Author(s): Kanki, T.
  • Journal Title: Mol.Cell.Biol. 24 Pages: 9823-9834
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Mitochondrial nucleoid and transcription factor A (2004)
  • Author(s): Kanki, T.
  • Journal Title: Ann N Y Acad Sci 1011 Pages: 61-68
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Physical and functional interaction between BH3-only protein Hrk and mitochondrial pore-forming protein p32 (2004)
  • Author(s): Sunayama, J.
  • Journal Title: Cell Death Differ 11 Pages: 771-778
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] High-Sensitivity Detection of the A3243G Mutation of Mitochondrial DNA (2004)
  • Author(s): Urata, M.
  • Journal Title: Clin Chem 50 Pages: 2045-2051
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Architectural role of TFAM in maintenance of human mitochondrial DNA. (2004)
  • Author(s): Kanki, T., Ohgaki, K., Gaspari, M., Gustafsson, C.M., Fukuoh, A., Sasaki, N., Hamasaki, N., Kang, D.
  • Journal Title: Mol.Cell.Biol. 24 Pages: 9823-9834
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Mitochondrial nucleoid and transcription factor A. (2004)
  • Author(s): Kanki, T., Nakayama, H., Sasaki, N., Takio, K., Alam, T.I., Hamasaki, N., Kang, D.
  • Journal Title: Ann N Y Acad Sci 1011 Pages: 61-68
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Physical and functional interaction between BH3-only protein Hrk and mitochondrial pore-forming protein p32. (2004)
  • Author(s): Sunayama, J., Ando, Y, Itoh, N., Tomiyama, A, Sakurada, K., Sugiyama, A., Kang, D., Tashiro, F., Gotoh, Y, Kuchino, Y, Kitanaka, C.
  • Journal Title: Cell Death Differ 11 Pages: 771-781
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] High-Sensitivity Detection of the A3243G Mutation of Mitochondrial DNA by a Combination of Allele-Specific PCR and Peptide Nucleic Acid-Directed PCR Clamping. (2004)
  • Author(s): Urata, M., Wada, Y., Kim, S., Chumpia, W., Kayamori, Y., Hamasaki, N., Kang, D.
  • Journal Title: Clin Chem 50 Pages: 2045-2051
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Physical and functional interaction between BH3-only protein HrK and mitochondrial pore-forming protein p32 (2004)
  • Author(s): Sunayama, J.
  • Journal Title: Cell Death Differ 11 Pages: 771-781
• [Publications] Chumpia, W.et al.: "Affinity purification of antibodies by using Ni2+-resins on which inclusion body-forming proteins are immobilized"Protein Expr Purif. 32. 147-150 (2003)
• [Publications] Yoshida, Y.et al.: "p53 Physically Interacts with Mitochondrial Transcription Factor A and Differentially Regulates Binding to Damaged DNA"Cancer Res.. 63. 3729-3734 (2003)
• [Publications] Okamaoto, M.et al.: "Ditercalinium chloride, a pro-anticancer drug, intimately associates with mammalian mitochondrial DNA and inhibits its replication"Curr.Genet.. 43. 364-370 (2003)
• [Publications] Suematsu, N.et al.: "Oxidative stress mediates tumor necrosis factor-alpha-induced mitochondrial DNA damage and dysfunction in cardiac myocytes"Circulation. 107. 1418-1428 (2003)
• [Publications] Alam, T.I.et al.: "Human mitochondrial DNA is packaged with TFAM"Nucleic Acids Res. 31. 1640-1645 (2003)
• [Publications] Kang, D., Hamasaki, N.: "Mitochondrial oxidative stress and mitochondrial DNA"CLin.Chem.Lab.Med.. 41. 1281-1288 (2003)