Molecular basis of ATP synthesis machinery from parasite living under microenvironment
Project/Area Number |
16K19114
|
Research Category |
Grant-in-Aid for Young Scientists (B)
|
Allocation Type | Multi-year Fund |
Research Field |
Parasitology (including sanitary zoology)
|
Research Institution | Nagasaki University |
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)
|
Project Period (FY) |
2016-04-01 – 2018-03-31
|
Project Status |
Completed (Fiscal Year 2017)
|
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)
|
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 |
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.
|
Report
(3 results)
Research Products
(27 results)
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
[Presentation] Glycosomal metabolic enzyme Isocitrate Dehydrogenase from Trypanosoma brucei shows dual coenzyme specificity2017
Author(s)
Xinying Wang, Daniel Ken Inaoka, Emmanuel O. Balogun, Fumie Hamano, Nicole Ziebart, Stefan Allmann, Michael Boshart, Frederic Bringaud, Fuyuki Tokumasu, Tomoo Shiba, Shigeharu Harada and Kiyoshi Kita
Organizer
第 77 回日本寄生虫学会東日本支部大会
Related Report
-
-
-
-
-
-