Investigation of new zinc finger protein function as a redox sensor for reactive oxygen in the cells

• Project/Area Number: 26350962
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Biomolecular chemistry
• Research Institution: Doshisha Women's College of Liberal Arts
• Principal Investigator: NEGI SHIGERU 同志社女子大学, 薬学部, 准教授 (50378866)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2016: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2014: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: 亜鉛フィンガータンパク質 / レドックス反応 / レドックスセンサー / DAN結合 / 過酸化水素 / 亜鉛フィンガー / レドックス / 酸化 / 転写因子 / レドックスレンサー / 金属結合

Outline of Final Research Achievements

In this study, we tried to investigate the oxidation reactivity, stability of the folding structure, DNA-binding property using Sp1 zinc finger protein.
As a result of an oxidation reaction with H2O2, reactivity became the following order, F1>>F2>F3, indicating that each three finger has a non-equivalence reactivity for the oxidation reaction. In DNA-binding assay using the gel shift method, oxidized Sp1 zinc finger protein lost the DNA-binding ability due to Zn(II) ion release following S-S bond formation.

Research Products

• [Journal Article] Intrinsic cell permeability of the GAGA zinc finger protein into HeLa (2015)
  • Author(s): Shigeru Neg*i, Yuka terada, Misato Suzuyama, Makoto Matsumoto, Akino Honbo, Yoko Amagase, Yumiko Mizukawa, Akiko Kiriyama, Kastumi Iga, Tetsuro Urushidani, Yukio Sugiura
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 464 Issue: 4 Pages: 1034-1039
  • DOI: 10.1016/j.bbrc.2015.07.060
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Efficient cleavage of DNA oligonucleotides by a non-FokI-type zinc (2015)
  • Author(s): Shigeru Negi*, Michiko Yoshioka, Hiroko Mima, Makoto Mastumoto, Michiko Suzuki, Mao Yokoyama, Koji Kano, Yukio Sugiura,
  • Journal Title: Bioorganic & Medicinal Chemistry Letter Volume: 25 Issue: 19 Pages: 4074-4077
  • DOI: 10.1016/j.bmcl.2015.08.045
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Supramolecular intracellular delivery of an anionic porphyrin by octaarginine-conjugated per-O-methyl-β-cyclodextrin (2015)
  • Author(s): H.Kitagisshi, F.Chai, S.Negi. Y.Sugiuraand K.Kano
  • Journal Title: Chem.Commun. Volume: 51 Issue: 12 Pages: 2421-2424
  • DOI: 10.1039/c4cc09042j
  • Peer Reviewed / Open Access
• [Presentation] Control of DNA-binding of GAL4 zinc finger by porphyrin-cyclodextrin supramolecular system (2014)
  • Author(s): Shigeru Negi、Koji Kano, Yukio Sugiura
  • Organizer: ICCP-8
  • Place of Presentation: Istanbul(Turkey)
  • Year and Date: 2014-06-22 – 2014-06-27
• [Presentation] Sp1亜鉛フィンガーの各フィンガードメインにおけるレドックス応答の非対称性 (2014)
  • Author(s): 根木 滋、福田あずさ、鎌手裕美香、杉浦幸雄
  • Organizer: 第41回生体分子科学討論会
  • Place of Presentation: 九州大学西新プラザ(福岡市）
  • Year and Date: 2014-06-06 – 2014-06-07
• [Presentation] 超分子ヘムタンパク質モデル化合物による生体内ＣＯの除去 (2014)
  • Author(s): 北岸宏亮、根木 滋、勇村亞希、杉浦幸雄、加納航治
  • Organizer: 第41回生体分子科学討論会
  • Place of Presentation: 九州大学西新プラザ(福岡市）
  • Year and Date: 2014-06-06 – 2014-06-07