| Project/Area Number |
16550147
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Chemistry related to living body
|
|---|
| Research Institution | Tokyo University of Pharmacy and Life Sciences |
|---|
Principal Investigator |
WATANABE Kinzo Tokyo University of Pharmacy and Life Sciences, School of Life Sciences, Associate Professor, 生命科学部, 助教授 (10167116)
|
|---|
| Project Period (FY) |
2004 – 2005
|
| Project Status |
Completed (Fiscal Year 2005)
|
| Budget Amount *help |
¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 2005: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2004: ¥1,800,000 (Direct Cost: ¥1,800,000)
|
|---|
| Keywords | prostaglandin / biosynthetic pathway / soft coral / Clavularia viridis / lipoxygenase / アラキドン酸カスケード / リポキシゲナーゼ / Clavularia virides / PCR / DNA塩基配列解析 / プロスタノイド |
|---|
| Research Abstract |
The Okinawan soft coral Clavularia viridis is a rich natural source of anti-tumor prostaglandins, clavulones. Corey et al. suggested that the prostaglandin biosynthesis of clavulones could occur through 8-lipoxygenase (8-LOX) pathway. In this study, the biosynthetic intermediates from arachidonic acid using an acetone powder from the living coral, and detected 8-hydroperoxyeicosatetraenoic acid and preclavulone-A were investigated. These biosynthetic intermediates were characterized by HPLC and NMR, and were found to be each enantiomeric in a ratio of 3:1 and 1:1, respectively. cDNA for the key enzyme of the 8-LOX pathway was isolated using degenerate origonucleotides which were designed based on conserved amino acid sequences of LOX proteins. The obtained cDNA encodes a protein of 1066 amino acids, which include an allene oxide synthase (AOS) domain in addition to a LOX domain. The deduced amino acid sequence shares high similarity to that of AOS-8R-LOX fusion protein from the Caribbean soft coral, Plexaura homomalla, cDNA of which was cloned by Brash et al. These results suggest a legitimacy of the proposed Corey's biosynthetic pathway.
|
