2021 Fiscal Year Research-status Report
赤痢アメーバをモデルとした寄生虫ポリアミン生合成経路の新展開
| Project/Area Number |
21K15427
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| Research Institution | The University of Tokyo |
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Principal Investigator |
Jeelani Ghulam 東京大学, 大学院医学系研究科(医学部), 助教 (60468519)
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| Project Period (FY) |
2021-04-01 – 2024-03-31
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| Keywords | Entamoeba histolytica / Polyamine / Protozoan parasite / Metabolisms / Drug developement |
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| Outline of Annual Research Achievements |
Expression and purification of two enzyme involved in polyamine biosynthesis pathway have been completed. Gene silencing of one polyamine biosynthesis pathway enzyme (Ornithine decarboxylase) in Entamoeba histolytica have been completed and some phenotypes (growth kinetics, cell shape) have been observed. Metabolome analysis of gene silencing transformant have been completed. Optimization of protein purification using hydrophobic (Butyl-Toyopearl) and anion exchange (MonoQ) columns have been established. A review article was submitted and accepted by Trends in parasitology (https://doi.org/10.1016/j.pt.2022.02.004)
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
Optimization of enzymatic assays (spermidine synthase, spermine synthase and others) using HPLC is taking time. Silencing of one of the novel polyamine pathway enzyme (annotated as ornithine/arginine/lysine decarboxylase in genome database) failed suggesting essential enzyme for parasite survival.
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| Strategy for Future Research Activity |
Complete functional characterization of the two enzyme which we expressed and identification of the there novel enzymatic activity.
Fluxomic based metabolomic approach in order to elucidate the whole polyamine metabolic pathway.
Purification of native E. histolytica polyamine pathway enzymes.
Transcriptional silencing and metabolome analysis of the identified enzyme and their phenotypic analysis.
Complete and submit manuscript for publication.
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| Causes of Carryover |
Due to the coronavirus pandemic, the planned use of the budget for conference attendance was not used.
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