| Project/Area Number |
21K14746
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| Research Category |
Grant-in-Aid for Early-Career Scientists
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| Allocation Type | Multi-year Fund |
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| Review Section |
Basic Section 37020:Chemistry and chemical methodology of biomolecules-related
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| Research Institution | Tokyo Institute of Technology |
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Principal Investigator |
ジャー トニーズィ 東京工業大学, 地球生命研究所, 特任助教 (10800328)
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| Project Period (FY) |
2021-04-01 – 2026-03-31
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| Project Status |
Granted (Fiscal Year 2021)
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| Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2022: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2021: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
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| Keywords | Origins of Life / Panspermia / Prebiotic Chemistry / Polyesters / LLPS / Protocell / Astrobiology / Biopolymers / Phase Separation |
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| Outline of Research at the Start |
Membraneless droplets generated by liquid-liquid phase separation (LLPS) were likely protocells. However, their structures and functions are unclear. We propose to study assembly, structure, and function of membraneless protocell compartments such as polyester droplets and liquid crystal coacervates. By probing the ability of LLPS protocells to stably compartmentalize and protect biomolecules and connecting these to essential modern cellular functions, we hope to understand more about how primitive protocells and chemistries directly evolved into modern cells and biologies.
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| Outline of Annual Research Achievements |
First, we explored the affect of temperature and chirality on polyester synthesis and droplet assembly from dehydration of various alpha hydroxyacid (AHA) monomer solutions. We found no significant difference in these aspects for the AHA monomers we tested, but there could be differences in other monomers, which we are currently testing. Next, we found that increasing temperature results in more propensity for polyester droplets degrade more readily, suggesting that protection of the internal components from heat will be limited by the stability of the droplets themselves.
We then were able to incorporate anionic AHAs into polyesters (previously not possible), which subsequently allowed us to observe the distribution of salts (sodium, calcium, and magnesium) segregating into the droplets through MS and micro-raman imaging. This suggests that different salts are compartmentalized within the membraneless protocells in a different manner depending on the salt and the protocell. In fact, interactions between one polyester sample and calcium results in crystallization, and simultaneous droplet and crystal existence may be indicative of primitive biomineralization. Finally, we are doing pilot studies to incorporate both lipids and lipid bilayers into/around the droplets, further spatial chemical analyses, and also incorporation of mineral nanoparticles. The next step is to test their function and compartmentalization stability within the droplets.
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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
I believe that the current project has been proceeding quite smoothly, despite some restrictions due to COVID-19. This has been supported by research and staff members in the PI's lab, as well as collaborators both inside Japan and internationally.
The experimental progress at ELSI and some other local collaborator labs has been unhindered. However, we have some planned analyses at other collaborative labs (both in Japan and Internationally) which were not possible due to travel restrictions for most of 2021. However, we are planning to finish these analyses soon, and hope to be able to visit those collaborator labs in this fiscal year.
Unfortunately, dissemination of the research at in-person meetings was also not possible, and thus we could only present the results at in-person conferences. However, I was able to attend many in-person conferences in FY2021, both internationally and domestically, and so we hope that others are aware of our progress.
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| Strategy for Future Research Activity |
First, we plan to finish our chemical analyses of the current systems synthesized and assembled at different temperatures and chiralities, and publish the manuscript in this fiscal year, which includes some discussion on the thermodynamics of polyester droplet assembly/stability and protection of analytes. Then, we hope to finish some physical analyses to prepare and publish a paper on biomineralization and crystallization of polyester microdroplets catalyzed by calcium salts.
Next, we will finish chemical and physical analyses (at collaborators' labs) to determine the spatial compartmentalization pattern of different salts in different polyester droplet types. This information will reveal which salts and which polyesters are more amenable to salt compartmentalization, and in turn, protection. These studies will tie into current collaborative studies with researchers studying vesicles and lipids, so that perhaps lipid vesicles can provide another boundary to increase the segregative and compartmentalization functions of polyester droplets. Depending on the progress of these studies, we may prepare another paper in FY2021. Finally, we will then move on to further probing of polyester droplets and their ability to compartmentalize and protect mineral nanoparticles as well as biomolecules such as peptides, proteins, and nucleic acids.
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