| Project/Area Number |
20370043
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Structural biochemistry
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| Research Institution | National Institute of Advanced Industrial Science and Technology |
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Principal Investigator |
SENDA Toshiya National Institute of Advanced Industrial Science and Technology, バイオメディシナル情報研究センター, 主任研究員 (30272868)
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| Co-Investigator(Renkei-kenkyūsha) |
HORIKOSHI Masami 東京大学, 分子細胞生物学研究所, 准教授 (70242089)
ENOMOTO Takemi 武蔵野大学, 薬学研究所, 教授 (80107383)
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| Project Period (FY) |
2008 – 2010
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| Project Status |
Completed (Fiscal Year 2010)
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| Budget Amount *help |
¥18,590,000 (Direct Cost: ¥14,300,000、Indirect Cost: ¥4,290,000)
Fiscal Year 2010: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2009: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000)
Fiscal Year 2008: ¥8,710,000 (Direct Cost: ¥6,700,000、Indirect Cost: ¥2,010,000)
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| Keywords | 結晶構造解析 / 結晶化 / ヒストン / ヌクレオソーム / ヒストンシャペロン / 精製 / FACT / 動的光散乱 / 大量発現 / X線結晶構造解析 / ヌクレオソーム構造変換 / 大量発現系 |
|---|
| Research Abstract |
FACT, which interacts with the histone H2A-H2B complex, is known to be involved in nucleosome assembly/disassembly. In order to reveal the molecular mechanism of nucleosome assembly/disassembly, crystal structure analysis of FACT was initiated. In the present study, we have established a method that can give about 7 mg of highly purified FACT from 1L culture of insect cell SF+. In addition, a purification method of histone proteins was improved. We confirmed that the purified FACT interacts with the histone proteins. To establish a proper condition for preparing the FACT-histone complex without forming aggregates, solution conditions of the FACT-histone complex were investigated by the dynamic light scattering method. On the basis these results, we are making an effort to obtain crystals of the FACT-histone complex.
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