| Project/Area Number |
23570155
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Structural biochemistry
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| Research Institution | National Cardiovascular Center Research Institute |
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Principal Investigator |
AKIYAMA Masashi 独立行政法人国立循環器病研究センター, 研究所, 室長 (30298179)
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| Co-Investigator(Kenkyū-buntansha) |
KOKAME Koichi 独立行政法人国立循環器病研究センター, 研究所, 室長 (40270730)
TAKEDA Soichi 独立行政法人国立循環器病研究センター, 研究所, 室長 (80332279)
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| Project Period (FY) |
2011 – 2013
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| Project Status |
Completed (Fiscal Year 2013)
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| Budget Amount *help |
¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2012: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2011: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
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| Keywords | ADAMTS13 / von Willebrand因子 / 結晶構造 / 酵素動力学 / VWF / ずり応力 / プロテインS |
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| Research Abstract |
A P475S polymorphism in the ADAMTS-13 gene causes an reduction in plasma VWF-cleaving activity. To demonstrate the impact of this mutation, we determined the crystal structure of the P475S mutant of ADAMTS13-DTCS (DTCS-P475S) and the enzymatic parameters of MDTCS-P475S. Structural analysis showed that the conformation of the V-loop was significantly different in DTCS-P475S, where no obvious interactions of Ser475 with other residues were observed. MDTCS-P475S showed a reaction rate similar to that of wild-type MDTCS, but showed two-fold lower affinity for the peptidyl substrate. MDTCS-P475S can moderately cleave shear-treated VWF. We have provided structural evidence that the P475S polymorphism in ADAMTS-13 leads to increased local structural instability, resulting in lowered affinity for the substrate without changing the reaction rate. The moderate activity of ADAMTS-13-P475S for shear-treated VWF is sufficient to prevent thrombotic thrombocytopenic purpura onset.
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