2013 Fiscal Year Final Research Report
Studies on the enzyme that catalyzes D/L-isomerization of an amino acid during the post-translational modification of neuropeptide precursor.
| Project/Area Number |
23570094
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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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| Research Field |
Animal physiology/Animal behavior
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| Research Institution | Hiroshima University |
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Principal Investigator |
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| Project Period (FY) |
2011 – 2013
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| Keywords | 神経科学 / 生理活性物質 / 細胞培養 / 遺伝子導入 / アメフラシ |
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| Research Abstract |
NdWFamide (NdWFa) is a D-amino acid containing neuropeptide of the marine gastropod, Aplysia kurodai. In this study, we attempt to identify the peptidyl D/L-isomerase (PI) that catalyzes the D-/L-conversion of NdWFa in Aplysia. To this end, we prepared NdWFa-expressing neuron specific cDNAs by the subtraction method. When the cDNAs were cloned and sequenced, we found that several predicted proteins with N-terminal signal peptide could not be specified their functions by homology search on database. We currently assume that those proteins are candidates for the PI of Aplysia.
Then, we tried to establish NdWFa-expressing cell-line, because such cells could be used for the bioassay system for the candidate genes of the PI. Transformation of NdWFa-precursor gene to CHO-K1 cell seems to be successful. However, biosynthesis of the peptide could not be confirmed, so far. We may need fine-tuning of experimental conditions that leads to more effective expression of foreign gene.
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