Structural Analysis of Ethylene (Plant Hormone) Sensor Protein

乙烯（植物激素）传感器蛋白的结构分析

• 批准号: 12680666
• 负责人: SHIRO Yoshitsugu
• 金额: $ 2.24万
• 依托单位: The Institute of Physical and Chemical Research
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12680666/
• 关键词: Ethylene; Plant Hormone; Two-component system; Histidine kinase; Copper (I) complex; XAFS; Sensor Protein; センサータンパク質; エチレン受容体; 二成分情報伝達系; リン酸移転; 銅

Ethylene is a plant hormone, which is significantly involved in life cycle of plants. Ethylene exhibits its physiological activities by virtue of binding to its sensor protein, ETR1. ETR1 belongs to the two-component regulatory system in cells, and mainly consists of two functional domain, sensor domain and histidine kinase domain. Most recently, it has been found that the ETR1 sensor domain contains Cu(I). In the present project,we have been trying to obtain the direct evidence that Cu(I) is the ethylene binding site of ETR1. We constructed expression system of the recombinant protein of the ETR1 sensor domain (trans-membrane protein) in E. coli, established it purification procedure, and measured the Cu K-edge XAFS spectra. The spectra is significantly different in the presence and absence of ethylene, showing that ethylene bind to the Cu(I) sitel of ETR1 and causes its coordination structural changes.

乙烯是一种植物激素，它与植物生命周期显着有关。乙烯通过与其传感器蛋白ETR1结合而表现出其生理活性。 ETR1属于细胞中的两个组分调节系统，主要由两个功能域，传感器结构域和组氨酸激酶结构域组成。最近，已经发现ETR1传感器域含有Cu（i）。在本项目中，我们一直在尝试获得直接证据，即Cu（i）是ETR1的乙烯结合位点。我们在大肠杆菌中构建了ETR1传感器结构域（跨膜蛋白）的重组蛋白的表达系统，确定了IT纯化程序，并测量了Cu k-gede XAFS光谱。在存在和不存在乙烯的情况下，光谱显着差异，表明乙烯与ETR1的Cu（I）结合并引起其协调性结构变化。

项目成果

K. Sakai,Y. Matsui, T. Kouyama, Y. Shiro and S.Adachi: "Optical monitoring of freeze-trapped reaction intermediates in protein crystals: a microspectrophotometer for cryogenic protein crystallography"J. Appl. Cryst. 35. 270-273 (2002)

K.酒井，Y.

Miyatake,H.,Mukai,M.,Park,S-Y.,Adachi,S.,Tamura,K., et al: "Sensory Mechanism of Oxygen Sensor FixL from Rhizobium meliloti : Crystallographic Mutagenesis and Resonance Raman Spectroscopic Studies"J.Mol.Biol.. 301. 415-431 (2000)

Miyatake,H.、Mukai,M.、Park,S-Y.、Adachi,S.、Tamura,K.等人：“苜蓿根瘤菌氧传感器 FixL 的感觉机制：晶体诱变和共振拉曼光谱研究”J.Mol

H.Kumita, S.Yamada, H.Nakamura, Y.Shiro: "Chimeric sensory kinases containing O2 sensor domain of FixL, and histidine kinase domain from thermophile"Biochem.Biophys.Acta. (印刷中). (2003)

H.Kumita、S.Yamada、H.Nakamura、Y.Shiro：“含有 FixL 的 O2 传感器结构域和来自嗜热菌的组氨酸激酶结构域的嵌合感觉激酶”Biochem.Biophys.Acta（出版中）。

D.-S.Lee, A.Yamada, H.Sugimoto, I.Matsunaga, H.Ogura, K.Ichihara, S.Adachi, S.-Y.Park, Y.Shiro: "Substrate Recognition and Molecular Mechanism of Fatty Acid Hydroxylation by Cytchrome P450 from Bacillus subtilis : Crystallographic, Spectroscopic and Mutat

D.-S.Lee、A.Yamada、H.Sugimoto、I.Matsunaga、H.Ogura、K.Ichihara、S.Adachi、S.-Y.Park、Y.Shiro：“脂肪的底物识别和分子机制

中村寛夫: "生物物理"シグナル変換2成分系タンパク質. 4 (2001)

Hiroo Nakamura：“生物物理学”信号转导双组分系统蛋白 4 (2001)。