| Budget Amount *help |
¥4,110,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥510,000)
Fiscal Year 2007: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2006: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Research Abstract |
Amyloid is extracellular deposit of insoluble fibrillar aggregate of proteins those are normally soluble in their native conformation, resulting in the impairment of tissue structure and function. We have established several amyloid fibril formation systems in vitro, including β-amyloid of Alzheimer's disease (Aβ) and β2-microglobulin (β2-m) amyloid (Aβ2M) of dialysis-related amyloidosis. Using these systems, important roles of various biological molecules or compounds on the formation and dissociation of amyloid fibrils were discovered. To model the amyloid fibril formation in vivo by in vitro experimental system or numerical model, we selected several phases of fibril formation or dissociation, and clarified their mechanism by biophysical, kinetic and thermodynamic analyses.
1. To understand the molecular pathogenesis of Aβ2M amyloidosis, the biological molecules that induce amyloid fibril formation by affecting the conformation and stability of β2-m and amyloid fibrils need to be ide
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ntified. We showed that anionic amphipathic compounds including non-esterified fatty acids (NEFAs) and some lysophospholipids induce the fibril extension at a neutral pH. Furthermore, hemodialysis patients had significantly higher plasma concentrations of lysophospholipids than healthy subjects. These results suggest possible role of lysophospholipids and NEFAs in the development of Aβ2M amyloidosis.
2. Using spectrophotometry, spectrofluorometry and surface plasmon resonance, we investigated the anti-amyloidogenic effects of five flavonoids on β-amyloid fibril (fAβ) in vitro. The results suggest that flavonoids, especially myricetin exert an anti-amyloidogenic effect in vitro by preferentially and reversibly binding to the amyloid fibril structure of fAβ, rather than to Aβ monomers.
3. We developed a high-throughput screening system for the nucleation of Aβ2M and Aβ amyloid fibril using thioflavin T fluorescence spectroscopy. Using this system, effect of various biological molecules including proteoglycans, glycosaminoglycans, were investigated. As the conclusion of this study, the remarkable advance in the clarification of the mechanism of Aβ2M and Aβ amyloid fibril formation were obtained. Less
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