Search for novel target molecules of intracellular amyloid beta

2016 Fiscal Year Final Research Report

• Project/Area Number: 15K12744
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Biomolecular chemistry
• Research Institution: Kyoto University
• Principal Investigator: Murakami Kazuma 京都大学, 農学研究科, 准教授 (80571281)
• Co-Investigator(Renkei-kenkyūsha): IZUMI Shunsuke 広島大学, 大学院理学研究科, 教授 (90203116) ; KUME Toshiaki 京都大学, 大学院薬学研究科, 准教授 (10303843)
• Research Collaborator: TOKUDA Maki ; KATAOKA Shotaro
• Project Period (FY): 2015-04-01 – 2017-03-31
• Keywords: アルツハイマー病 / アミロイドβ / オリゴマー / 標的探索 / MS/MS / シグナル伝達 / ビオチン化 / 磁性ビーズ

Outline of Final Research Achievements

The 42-mer amyloid β-protein (Aβ42) oligomerizes (2~24-mer: 2 or 3 x n-mer) intracellularly to show the neurotoxicity in the pathogenesis of Alzheimer′s disease. There are few studies on the mode of action of intracellular toxic oligomers of Aβ42 and their target molecules. We developed a molecular probe of toxic oligomers of Aβ42 and searched for their associated proteins within the cell. The design of the probe was based on the toxic-conformation constrained E22P-Aβ42 and L,L-2,6-diaminopimeric acid (DAP) linker at position 40 in the C-terminal hydrophobic core, the formation of which plays a critical role in the oligomerization of Aβ42. The dimer probe of Aβ42 generated oligomers (12~24-mer) with the potent neurotoxicity against human SH-SY5Y neuroblastoma cells. After reacting the probe with cytosolic lysates of SH-SY5Y, three proteins preferably bound to the dimer probe were identified by MS/MS analysis using the biotin-avidin technology.

Free Research Field

生物有機化学