| Project/Area Number |
23310160
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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 |
Chemical biology
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| Research Institution | Kyoto University |
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Principal Investigator |
HATTORI Akira 京都大学, 薬学研究科(研究院), 准教授 (50300893)
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| Co-Investigator(Kenkyū-buntansha) |
OISHI Shinya 京都大学, 薬学研究科, 講師 (80381739)
FUJIWARA Hiroshi 金沢大学, 医学研究科, 教授 (30252456)
INOUE Hideshi 東京薬科大学, 生命科学部, 教授 (20184765)
MARUYAMA Masato 関西医科大学, 医学部, 講師 (00399445)
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| Co-Investigator(Renkei-kenkyūsha) |
NISHIMURA Shinichi 京都大学, 薬学研究科, 助教 (30415260)
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| Project Period (FY) |
2011-04-01 – 2014-03-31
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| Project Status |
Completed (Fiscal Year 2013)
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| Budget Amount *help |
¥20,020,000 (Direct Cost: ¥15,400,000、Indirect Cost: ¥4,620,000)
Fiscal Year 2013: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2012: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2011: ¥9,620,000 (Direct Cost: ¥7,400,000、Indirect Cost: ¥2,220,000)
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| Keywords | ユビキチン / 脱ユビキチン化酵素 / activity-based probe / 活性制御 / 酸化ストレス / プローブ / ユビキチン様ドメイン / プロテアーゼ / アクティビティーベースドプローブ / プロテオミクス解析 / activety-based probe |
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| Research Abstract |
Post-translational modification of cellular proteins by ubiquitin (Ub) is involved in various aspects of cell physiology, such as protein degradation via proteasome, DNA repair, and membrane trafficking. Deubiqutinating enzymes liberate a Ub moiety from polyUb chains attached on substrate proteins. In the current study, we investigated novel research tools for the enzymatic characterization of deubiquitinating enzymes. Ub-granzyme B (Ub-GrB), an N-terminal Ub fusion mature granzyme B was expressed in a baculovirus system. By employing Ub-GrB, we showed that human ubiquitin specific protease 47 is active. In addition, we explored oxidative stress-sensitive deubiquitinating enzymes, and found that ubiquitin C-terminal hydrolase-L3 is susceptible to reactive oxygen species by utilizing a Ub activity-based probe, Ub-vinyl sulfone.
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