Molecular Design and Highly Efficient Synthesis of Molecular Probes Useful for Investigation of Molecular Recognition Mechanisms of Ligand-Receptor Interaction in Gibberellins

用于研究赤霉素配体-受体相互作用的分子识别机制的分子探针的分子设计和高效合成

• 批准号: 09672183
• 负责人: SETO Hideharu
• 金额: $ 1.98万
• 依托单位: The Institute of Physical and Chemical Research (RIKEN)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09672183/
• 关键词: gibberellin; receptor; photoaffinity-labeling; fluorescein-labeling; α-amylase; synthesis; brassinosteroid; jasmonic acid; three hybrid system; アリューロン細胞

This study intended to design and synthesize gibberellin (GA)-based molecular probes, fluorescein- and photoaffinity-labeled GAs, useful for investigation of molecular recognition mechanisms of ligand-receptor interaction in a phytohormone, GAs. In order to enhance the chemical stability of GA molecule and seek the position suitable for attaching the spacer-functional site, a variety of GA analogs were synthesized from gibberellic acid (GAィイD23ィエD2), and their biological activities were evaluated by α-amylase induction test using half cut barley seeds. On the basis of these data on the structure-activity relationships, fluorescein- and photoaffinity-labeled GAs were respectively assembled via 17-hydroxy-16β, 17-dihydro-19-deoxoGAィイD21ィエD2 derivative.An experiment on the interaction between the fluorescein-labeled GA with a pyrene moiety as the functional site and protoplasts isolated from the barley aleurone layers suggested that GA receptors might be located at the external face of the aleurone plasma membrane. The photoaffinity-labeled GA with aryl diazirine moiety as the functional site is expected to be used for isolation and identification of GA-binding proteins including GA-receptors.To apply the same synthetic strategies to assembling brassinosteroid (BR)- and jasmonic acid (JA)-based molecular probes, the synthetic modification of BR and JA were also carried out to accumulate the data concerning their structure-activity relationships. During the studies, deuterated-, 5-epi-, 6a-carba- and extra-hydroxylated-brassinolides were synthesized. In addition, a variety of JA derivatives including JA-dexamethasone hybrid ligand were synthesized.

本研究旨在设计和合成基于赤霉素（GA）的分子探针，荧光素和光亲和标记的GAs，用于研究植物激素GAs中配体-受体相互作用的分子识别机制。为了提高赤霉素分子的化学稳定性，寻找适合连接间隔子功能位点的位置，以赤霉素（GA）D23为原料合成了多种GA类似物，并以半切大麦种子为材料，通过α-淀粉酶诱导试验评价了其生物活性。在这些构效关系数据的基础上，分别通过17-羟基-16 β，17-dihydro-19-deoxoGA β-D21 β-D2衍生物。用芘标记的GA作为功能位点，从大麦糊粉层分离的原生质体表明GA受体可能位于糊粉层质膜的外表面。以芳基二氮杂环丙烯为功能位点的光亲和标记GA有望用于GA结合蛋白（包括GA受体）的分离和鉴定，为了将同样的合成策略应用于油菜素内酯（BR）和茉莉酸（JA）分子探针的组装，我们还对BR和JA进行了合成修饰，以积累它们的构效关系数据。在研究过程中，合成了氘代、5-表-、6a-碳-和外羟基-油菜素内酯。此外，还合成了多种JA衍生物，包括JA-地塞米松杂合配体。

项目成果

H.Seto, S.Fujioka, H.Koshino, T.Suenaga, S.Yoshida, T.Watanabe and S.Takatsuto: "Epimerization at C-5 of Brassinolide with Sodium Methoxide and the Biological Activity of 5-epi-Brassinolide in the Rice Lamina Inclination Test"J. Chem. Soc., Perkin Trans.1

H.Seto、S.Fujioka、H.Koshino、T.Suenaga、S.Yoshida、T.Watanabe 和 S.Takatsuto：“油菜素内酯在 C-5 处与甲醇钠的差向异构化以及 5-表位油菜素内酯的生物活性

Hideharu Seto: "2,3,5-Tri-epi-brassinolide : Preparation And Biological Activity in Rice Lamina Inclination Test"Phytochemistry. 52(5). 815-818 (1999)

Hideharu Seto：“2,3,5-Tri-epi-brassinolide：制备及其在水稻叶片倾斜试验中的生物活性”植物化学。

Hideharu Seto: "Reactivities of Gibberellins with a Cationic Exchange Resin, Dowex-50W"Heterocycles. 52(2)(in press). (2000)

Hideharu Seto：“赤霉素与阳离子交换树脂，Dowex-50W 的反应性”杂环。

H.Seto, S.Fujioka, H.Koshino, T.Suenaga, S.Yoshida, T.Watanabe and S.Takatsuto: "2,3,5-Tri-epi-brassinolide: Preparation And Biological Activity in Rice Lamina Inclination Test"Phytochemistry. 52 (5). 815-818 (1999)

H.Seto、S.Fujioka、H.Koshino、T.Suenaga、S.Yoshida、T.Watanabe 和 S.Takatsuto：“2,3,5-Tri-epi-brassinolide：水稻叶片倾斜试验中的制备和生物活性

Hideharu Seto: "A General Approach to Synthesis of Labeled Brassinosteroids:Preparation of [25,26,27-^2H_7]Brassinolide with 60% Isotopic Purity from the Parent Brassinolide." Tetrahedron Lett.39(41). 7525-7528 (1998)

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