| Project/Area Number |
16K19114
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Parasitology (including sanitary zoology)
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| Research Institution | Nagasaki University |
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Principal Investigator |
INAOKA Ken Daniel 長崎大学, 熱帯医学・グローバルヘルス研究科, 助教 (10623803)
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| Project Period (FY) |
2016-04-01 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2017)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2017: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Keywords | 寄生虫 / ATP合成 / ASCT / 生化学的解析 / 結晶構造解析 / ミトコンドリア様オルガネラ / 寄生虫学 / 感染症 / 酵素 / 蛋白質 / 生体分子 / 特殊環境 |
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| Outline of Final Research Achievements |
Trypanosoma brucei produces acetate (Ace) in mitochondria by two different enzymes: acetyl-CoA (ACoA) hydrolase or acetate:succinate CoA-transferase (ASCT). Under physiological condition, ASCT transfers the CoA from ACoA to succinate (Suc) to form Ace and succinyl-CoA (SCoA), which is consequently converted back to Suc by SCoA synthase (SCS) with net production of 1 mol of ATP per mol of ACoA in a process named ASCT/SCS cycle. So far, no biochemical information for ASCT family of CoA-transferase is reported. In this project, an over-expression system and purification of active TbASCT together with a novel sensitive method to measure ASCT activity have been developed. Purified TbASCT obeys Michaelis-Menten kinetics with Kcat of 202 ATP per second and show high affinity for ACoA (Km = 47 μM) than succinate (9.86 mM). A method for in-gel ASCT activity staining was developed showing evidence that TbASCT is active as homotetramer which was consistent to the crystal structure.
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