| Project/Area Number |
23K05713
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 43040:Biophysics-related
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| Research Institution | Kanazawa University |
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Principal Investigator |
NGO XUANKIEN 金沢大学, ナノ生命科学研究所, 特任助教 (60778190)
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| Project Period (FY) |
2023-04-01 – 2026-03-31
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| Project Status |
Granted (Fiscal Year 2023)
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| Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2025: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2024: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2023: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
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| Keywords | ABC transporter / MsbA / LptC / HS-AFM / E. coli / FRET / LptB2FG / High-speed AFM |
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| Outline of Research at the Start |
The applicant will unveil how ATP binding and hydrolysis in the NBDs of MsbA allosterically regulate conformational transitions in the TMDs and elucidate how MsbA, LptC, and LptB2FG synergistically work together for extracting and transporting lipid A/LPS from Inner Membrane toward Outer Membrane.
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| Outline of Annual Research Achievements |
1)Successfully prepared 2D crystal structures of MsbA within lipid membranes containing both with and without Kdo2 lipid A; 2) Achieved high spatial and temporal imaging of both NBD and TMD sides of MsbA embedded in these lipid membranes; 3) Developed new methods for the automatic analysis of AFM data; 4) completed the construction of plasmids and transfected E. coli cells with the target plasmids aimed for FRET measurements.
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| Current Status of Research Progress |
Current Status of Research Progress
1: Research has progressed more than it was originally planned.
Reason
I am potentially investigating a novel pathway for the gate operation of Lipopolysaccharide (LPS) induced by ABC transporters MsbA and LptC in E. coli.
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| Strategy for Future Research Activity |
In FY2024, my planned research agenda includes the following tasks: 1)Finalize the data analyses of AFM data; 2) Develop a protein-membrane model suitable for Molecular Dynamics simulation and/or Coarse-Grained simulation; 3) Investigate the interaction of MsbA with target protein in E. coli using FRET techniques; 4) Draft the manuscript detailing the findings.
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