Role of ceramide during apoptosis induction

• Project/Area Number: 26450291
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Aquatic life science
• Research Institution: Nihon University
• Principal Investigator: YABU Takeshi 日本大学, 生物資源科学部, 研究員 (00551756)
• Co-Investigator(Kenkyū-buntansha): 山下 倫明 国立研究開発法人水産研究・教育機構, 水産大学校, 教授 (80344323) ; 今村 伸太朗 国立研究開発法人水産研究・教育機構, 中央水産研究所, 主任研究員 (80510007) ; 司馬 肇 (張培淦 / 司馬 肇(張培淦)) 日本大学, 生物資源科学部, 教授 (90256834)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: セラミド / アポトーシス / セラミド結合性タンパク質 / 解離定数 / 代謝・酵素 / セラミド結合蛋白質 / 水産化学 / ストレス

Outline of Final Research Achievements

As a biochemical approach to identify direct targets of ceramide, a sphingolipid that plays an important role in cell signaling as a second messenger, we performed an affinity chromatography. Western blot analysis of the eluted proteins using a variety of antibodies against both pro- and anti-apoptotic molecules, we successfully identified the B-cell lymphoma interacting mediator (Bim) as a novel ceramide-binding molecule. Using the purified recombinant Bim protein and its truncated form expressed, we carried out a series of in vitro binding experiments. Our results indicated that ceramide specifically binds to Bim protein through a hydrophobic domain located at its C-terminal end; the dissociation constant was 21.71 nM. Our present study revealed that specific binding of ceramide to the C-terminal hydrophobic domain of Bim is a critical step for signaling the cells to apoptosis through the destruction of mitochondria in response to either cytokine stimulations or stress conditions.

Research Products

• [Journal Article] Effects of IFN<gamma> administration on allograft rejection in ginbuna crucian carp (2016)
  • Author(s): Shibasaki Y, C. Hatanaka, Y. Matsuura, R. Miyazawa, T. Yabu, T. Moritomo, T. Nakanishi
  • Journal Title: Dev. Comp. Immunol. Volume: 62 Pages: 108-115
  • DOI: 10.1016/j.dci.2016.04.021
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Sequential steps of macroautophagy and chaperone-mediated autophagy are involved in the irreversible process of posterior silk gland histolysis during metamorphosis of Bombyx mori. (2016)
  • Author(s): Shiba, H., T. Yabu, M. Sudayama, N. Mano, N. Arai, T. Nakanishi, and K. Hosono
  • Journal Title: J. Exp. Biol. Volume: 1 Pages: 1-10
  • DOI: 10.1242/jeb.130815
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Recombinant carp IL-4/13B stimulates in vitro proliferation of carp IgM+ B cells (2016)
  • Author(s): Takuya Yamaguchi, Shunsuke Miyata, Fumihiko Katakura, Takahiro Nagasawa, Yasuhiro Shibasaki, Takeshi Yabu, Uwe Fischer, Chihaya Nakayasu, Teruyuki Nakanishi, Tadaaki Moritomo
  • Journal Title: Fish & Shellfish Immunology Volume: 49 Pages: 225-229
  • DOI: 10.1016/j.fsi.2015.12.043
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Purification and characterization of a fish granzymeA involved in cell-mediated immunity. (2016)
  • Author(s): Matsuura Y, T. Yabu, H. Shiba, T. Moritomo, T. Nakanishi
  • Journal Title: Dev Comp Immunol. Volume: 60 Pages: 33-40
  • DOI: 10.1016/j.dci.2016.02.011
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Exploring erythropoiesis of common carp (Cyprinus carpio) using an in vitro colony assay in the presence of recombinant carp kit ligand A and erythropoietin (2015)
  • Author(s): Fumihiko Katakura, Takeshi Yabu, Takuya Yamaguchi, Jiro Miyamae, Yuki Shirinashihama, Teruyuki Nakanishi, Tadaaki Moritomo
  • Journal Title: Developmental and Comparative Immunology Volume: 53 Issue: 1 Pages: 13-22
  • DOI: 10.1016/j.dci.2015.06.006
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Kinetics of lymphocyte subpopulations in allogeneic grafted scales of ginbuna crucian carp (2015)
  • Author(s): Shibasaki Y., Matsuura Y., Toda H., Imabayashi N., Nishino T., Uzumaki K., Hatanaka C.,Yabu T., Moritomo T, NakanishiT.,
  • Journal Title: Dev Comp Immunol Volume: 52 Issue: 1 Pages: 75-80
  • DOI: 10.1016/j.dci.2015.04.013
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Stress-induced ceramide generation and apoptosis via the phosphorylation and activation of nSMase1 by JNK signaling. (2015)
  • Author(s): T. Yabu, H. Shiba, Y. Shibasaki, T. Nakanishi, S. Imamura, K. Touhata, M. Yamashita.
  • Journal Title: Cell Death Differ. Volume: 22 Pages: 258-273
  • Peer Reviewed / Open Access / Acknowledgement Compliant