Metabolic regulation of estrogen-synthesizing enzymes by post-translational modification
| Project/Area Number |
18591034
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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 |
Endocrinology
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| Research Institution | Fujita Health University |
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Principal Investigator |
HARADA Nobuhiro Fujita Health University, School of Medicine, Professor (00189705)
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| Co-Investigator(Kenkyū-buntansha) |
SASAKI Emi Fujita Health University, School of Medicine, Assistant Professor (20178635)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥4,010,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥510,000)
Fiscal Year 2007: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2006: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | Aromatase / Post-translational Modification / Protein Degradation / Proteasome / Ubiouitination / Chaperone / Phosphorylation / Estrogen / ステロイドサルファターゼ / 酵素誘導 / 代謝回転速度 / ユビキチンリガーゼ |
|---|
| Research Abstract |
In this study project, we focussed on the post-translational modification of aromatase, a rate-limiting enzyme of estrogen production, and analyzed molecular and regulatory mechanisms of the post-translational modification and physiological roles of this reaction. Western blotting analysis showed a possibility of post-translational modification of aromatase by phosphorylation in the cultured cells in the presence of various inducers and regulatory factors. Furthermore, ELISA assay of aromatase contents in the cultured cells suggested a possiblity that the intracellular degradation rate (the turnover rate) was modified by various factors. So, The in vitro assay system was introduced to analyze the process of the intracelluar degradation of the aromatase in the endoplasmic reticulum (microsomal fraction). The degradation of the microsomal aromatase content and activity required the presence of ATP-Mg2+ and the cytoplasmic fraction in the reaction. An aromatase antibody-immunoreactive protein with a high molecular weight (AR-ir protein) appeared in parallel with disappearance of the authentic aromatase with the molecular weight of 53, 000 by Western blotting analysis. The disappearance of the aromatase was inhibited by presence of Geldanamycin, an inhibitor of HSP90 chaperone protein. However, because the purified recombinant aromatase did not disappear and the AR-ir protein did not appear in the assay and also because the microsomal aromatase did not disappear when low concentration of cholic acid was added to the assay system to solubilize the microsomal membrane, it was suggested that the microsomal aromatase was quantitatively regulated by the microsomal membrane-dependent degradation system together with cytoplasmic regulatory proteins. We are now analyzing the constitutive factors participating in the microsomal membrane-dependent degradation by using FLAG-tagged aromatase.
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Report
(3 results)
Research Products
(12 results)
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[Journal Article] The characteristics of aromatase deficient hairless mice indicate important roles of extragonadal estrogen in the skin.2008
Author(s)
Tsukahara, K., Kakuo, S., Moriwaki, S., Hotta, M., Ohuchi, A., Kitahara, T., Harada, N.
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Journal Title
J. Steroid Biochem. Mol. Biol. 108
Pages: 82-90
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Journal Article] Concomitant treatment of severe uterine adenomyosis in a Premenopausal woman with an aromatase inhibitor and a gonadotropin-releasing hormone agonist.2007
Author(s)
Kimura, F., Takahashi, K., Takebayashi, K., Fujiwara, M., Kita, N., Noda, Y., Harada, N.
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Journal Title
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Mode of action and functional significance of estrogen-induced dendritic growth, spinogenesis, and synaptogenesis in the developing Purkinje cell.2007
Author(s)
Sasahara, K., Shikimi, H., Haraguchi, S., Sakamoto, H., Honda, S., Harada, N., Tsutsui, K.
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Journal Title
J. Neurosci 27
Pages: 7408-7414
Description
「研究成果報告書概要(欧文)」より
Related Report
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