| 研究課題/領域番号 |
16K08259
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| 研究機関 | 国立研究開発法人理化学研究所 |
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研究代表者 |
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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| 研究期間 (年度) |
2016-04-01 – 2019-03-31
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| キーワード | Phosphatidylglucoside / GPRC / Lipids / Biosynthesis / GPR55 |
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| 研究実績の概要 |
The specificity problem of the commercially available antibody has been solved by systematic characterization of its binding specificity.
The study of the receptor (GPR55) specificity towards lyso-phosphatidylglucoside (its native ligand) and synthetic analogues has been completed and the manuscript has been submitted to an international peer reviewed journal. To this end, synthetic access to lyso-phosphatidylglucoside and its ether and amide analogue has been established and the biological activity has been assessed utilizing in vitro axon turning assays. The result of the bio-assay correlated well with the results of the molecular dynamics simulation conducted in parallel. This allowed further characterization of the GPR55 ligand binding pocket and paves the way towards development of novel GPR55 agonist classes. Furthermore, the ligand entry port on the extracellular face of GPR55 has been characterized, a prerequisite for targeted molecular design towards specific GPR55 antagonists.
The mass spectroscopic method developed to detect lyso-phosphatidylglucoside has been adopted to study glycosyl-inositol-phospoho-ceramides from plants, leading to a publication in the journal Science.
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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
Problems experienced during last fiscal year concerning the low specificity of commercial antibodies have been overcome. This allowed the project to proceed as planned.
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| 今後の研究の推進方策 |
After completion of in vitro assay system optimization, precursor structural requirements and the general enzyme kinetics will be determined as planned. As the crystal structure of the target enzyme has recently been reported, we will perform docking studies to further characterize and rationalize the structure-activity relationship. In parallel, the enantiomeric purity of the artificial fluorescent precursor of lyso-phosphatidylglucoside for in vitro studies will be further improved by optimizing the synthetic procedures.
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