| 研究課題/領域番号 |
20K16233
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| 研究機関 | 東京大学 |
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研究代表者 |
サントス ハルベルト・ヒメネス 東京大学, 大学院医学系研究科(医学部), 助教 (90793779)
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| 研究期間 (年度) |
2020-04-01 – 2022-03-31
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| キーワード | membrane contact site / Entamoeba histolytica / mitosome / endosome |
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| 研究実績の概要 |
Imaging analyses (indirect fluorescence assay, immunoelectron microscopy, live imaging) and fractionation of proteins involved in mitosome endosome contact sites (MECSs) have been completed. Fractionation experiments have been completed and corroborate the imaging data. Immunoprecipitation to detect other interacting partners of MECS was also finished. Blue-Native PAGE was completed and protein complexes were determined and analyzed. Plasmids for BioID-based proteomics are also ready. Functional characterization including liposome-based assays are being optimized. Preliminary results were presented in the Japan Society of Parasitology held last April 16-17, 2021. A mini review article was submitted and accepted by Parasitology International on April 27, 2021.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
Optimization of liposome-based assays is taking time. Silencing of ETMP1 is not possible as it is essential to the parasite. Establishment of dominant-negative mutant for EHD1 is also challenging as the parasite growth is severely hampered.
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| 今後の研究の推進方策 |
1. Characterize some candidate interacting proteins found in MECSs.
2. Complete functional characterization by analyzing effect of overexpression or silencing MCS-related components on endocytic pathway-related processes (endocytosis, phagocytosis, exocytosis etc).
3. Assess the effect of protein overexpression or silencing on mitosome function (sulfate activation).
4. Test whether contact sites involving EHD1 enhance lipid transport by lipid transfer proteins by liposome-based assay.
5. Perform RNAseq analysis of transcriptionally-silenced strain(s).
6. Complete and submit manuscript for publication.
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| 次年度使用額が生じた理由 |
Due to the coronavirus pandemic, the planned use of the budget for conference attendance was not used.
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