Role of ceramide during apoptosis induction

2016 Fiscal Year Final Research Report

• 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)
• Project Period (FY): 2014-04-01 – 2017-03-31
• 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

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.

Free Research Field

水産化学　脂質生化学