2017 Fiscal Year Research-status Report
Study on the thermodynamics and regulation of amyloid formation using calorimetry
| Project/Area Number |
17K07362
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| Research Institution | Osaka University |
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Principal Investigator |
李 映昊 大阪大学, たんぱく質研究所, 講師 (70589431)
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| Project Period (FY) |
2017-04-01 – 2019-03-31
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| Keywords | Amyloid fibril / Calorimetry / Energy landscape / Kinetics / Thermodynamics / Protein aggregation / Protein misfolding / Misfolding and disease |
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| Outline of Annual Research Achievements |
To study thermodynamics of protein misfolding-induced aggregation and establish a new assay, isothermal titration calorimetry (ITC) was performed on the formation of amyloid fibrils of several amyloidogenic proteins and peptides. I revealed that amyloid formation is accompanied by exothermic heat after the lag time, and obtained thermodynamic parameters such as the change in enthalpy using in-house algorithms. I further challenged to elucidate the kinetic property of amyloid generation, and succeeded in obtaining the rate constant of nucleation and elongation of amyloid fibrillation. Energy landscape was also built using parameters obtained. Taken together, an ITC-based assay was demonstrated to be a markedly useful method for the thermodynamic and kinetic study of protein aggregation.
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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 could fully obtain satisfactory results based on thermodynamic and kinetic analyses. Especially, an algorithm for the accurate and precise ITC analysis was successfully made. A new assay using ITC was also firmly established.
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| Strategy for Future Research Activity |
In 2017, I was able to estalish an ITC assay including the way of the experiment and analysis in detail. Therefore, in 2018, more amyloidogenic precursor proteins and peptides which were not examined in 2017 will be investigated using the same methodologies. By doing this, I will aim at revealing a general property and mechanism of amyloid formation. Lastly, great efforts will be added to establish another new assay using ITC, which monitors the inhibition of amyloid generation as well as destruction of preformed amyloid fibrils.
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