| Project/Area Number |
17550085
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Analytical chemistry
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| Research Institution | Tokyo University of Pharmacy and Life Science |
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Principal Investigator |
TASHIRO Sakurako Tokyo University of Pharmacy and Life Science, School of Pharmacy, Lecturer (40328555)
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| Co-Investigator(Kenkyū-buntansha) |
KOJIMA Masaki Iwate Medical University, School of Pharmacy, Associate Professor (90277252)
TASHIRO Mitsuru Meisei University, Faculty of Science and Technology, Associate Professor (40315750)
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| Project Period (FY) |
2005 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,510,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥210,000)
Fiscal Year 2007: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2006: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2005: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | amyloid fibrils / Alzheimer's disease / familial Parkinson's disease / α-synuclein / intermediate / アミロイド / NACP |
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| Research Abstract |
NAC (Non-Aβ-Component of amyloid) was identified as one of the component in amyloid fibril for Alzheimer's disease during the purification process of amyloid fibrils. Later its precursor protein, α-synuclein was cloned. Moreover, the point mutations of this precursor protein (A30P, A53T, E46K) were identified in the confirmed familial cases of autosomal-dominantly inherited, early-onset Parkinson's disease(PD). Now α-synuclein is used as an marker protein for Lewy bodies in PD patient in order to diagnose the disease.
In this study, we have attempted to examine the structural changes during the early stages of amyloid formation and also the effects of point mutations mentioned above using thioflavin T fluorescence, CD, NMR, small angle X-ray scattering (SAXS), and electron microscopy in order to elucidate the fibrillation mechanisms at an atomic or molecular level.
SVD analysis of CD and SAXS data obtained during amyloid formation of α-synuclein has revealed the possible existence of intermediate during the early stage of amyloid formation. Also the analysis of 1H-15N HSQC spectra at the early stage of fibrillation indicated the decrease in peak intensities corresponding to KTKEGV motif. As the decrease in peak intensities could be correlated to increase in molecular weight, KTKEGV regions might be involved in the early stage of fibril formations. Furthermore, the detailed SVD analysis of SAXS data showed possibility of the early stage intermediate being heptamer.
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