Studies on the molecular mechanism of sphingolipid metabolism based on the crystal structure of the enzymes
| Project/Area Number |
21570149
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Functional biochemistry
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| Research Institution | Osaka Medical College |
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Principal Investigator |
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| Co-Investigator(Renkei-kenkyūsha) |
HAYASHI Hideyuki 大阪医科大学, 医学部, 教授 (00183913)
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| Project Period (FY) |
2009 – 2011
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| Project Status |
Completed (Fiscal Year 2011)
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| Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2011: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2010: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2009: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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| Keywords | 酵素の触媒機構 / タンパク質 / スフィンゴ脂質 / セリンパルミトイル転移酵素 / スフィンゴシン1-リン酸リアーゼ / 酵素反応 / 生体分子 / 微生物 / スフィンゴシン1-リン酸 |
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| Research Abstract |
The crystal structural of serine palmitoyltransferase complexed with L-serine was determined. The multifunctional role of the catalytic residue, histidine, in the reaction mechanism was experimentally proposed by the analyses of the mutant SPTs, as follows ; The histidine residue fixes the conformation of both of L-serine and reaction product preventing the unfavorable side reactions, and enhances the Claisen-type condensation and decarboxylation by its function as an acid catalyst. Sphingosine 1-phosphate was overproduced successfully in E. coli. Assay system using the purified recombinant enzyme and fluorescent labeled-substrate was developed and enzymatic property was analyzed in detail. Sphingosine 1-phosphate was also crystallized with good reproducibility.
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Report
(4 results)
Research Products
(43 results)
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[Journal Article] Role of a conserved arginine residue during catalysis in serine palmitoyltransferase.2011
Author(s)
Lowther, J, Charmier, G., Raman, M. C., Ikushiro, H., Hayashi, H., Campopiano, D. J.
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Journal Title
FEBS Lett.
Volume: Vol.585, No.12
Pages: 1729-1734
Related Report
Peer Reviewed
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[Journal Article] Structural insights into the enzymatic mechanism of serine palmitoyltransferase from Sphingobacterium multivorum.2009
Author(s)
Ikushiro, H., Islam, M. M., Okamoto, A., Hoseki, J., Murakawa, T., Fujii, S., Miyahara, I., Hayashi, H.
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Journal Title
J. Biochem.
Volume: Vol.146, No.4
Pages: 549-562
NAID
Related Report
Peer Reviewed
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