2016 Fiscal Year Research-status Report
First neuronal guidance lipid: Elucidation of biosynthesis and development of lead compounds to support neuronal recovery after injury
| Project/Area Number |
16K08259
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| Research Institution | Institute of Physical and Chemical Research |
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Principal Investigator |
GREIMEL PETER 国立研究開発法人理化学研究所, 脳科学総合研究センター, 専任研究員 (60525541)
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| Project Period (FY) |
2016-04-01 – 2019-03-31
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| Keywords | Phosphatidylglucoside / GPRC / Lipids / Biosynthesis |
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| Outline of Annual Research Achievements |
Due to delays in gaining access to cell-lines stably over-expressing the target bio-synthetic enzymes and thus in accessing purified target enzymes the planned in vitro assay series has been slightly delayed (see blow). In the meantime, the prepared reference material was used to probe receptor specificity in ex vivo assay systems, such as neuronal ex-plants. Additionally, a homology model of the target G-coupled protein receptor has been developed and is now ready for validation. Utilizing the established synthetic route via P(III) towards the target lipids, a novel, switch-able, fluorescent lipid analogue has been developed and its application for in vitro assay systems has been established. Publication of these results has been accepted in the Journal Scientific Reports.
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| Current Status of Research Progress |
Current Status of Research Progress
3: Progress in research has been slightly delayed.
Reason
An unexpected problem with low specificity of commercial antibodies against the target enzyme occurred. Extra time was required to identify the most potent antibody against the target enzyme, a pre-requisite to validate over-expression of target enzyme in cell lines for in vitro assay. Synthetic work towards acceptor lipids is progressing as expected.
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| Strategy for Future Research Activity |
The homology model established during the last fiscal year will be validated against the experimental ex vivo results of PtdGlc analogues utilizing molecular dynamics simulations of the receptor and synthetic agonist/antagonist in a membrane environment. This will allow to develop a structure activity map and provide the basis for the next publication. Experimentally, after gaining access to sufficient purified target enzyme, the in vitro assays will be performed as planned.
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