Study on the molecular basis for the regulation of redox metabolism network in plastids
质体氧化还原代谢网络调控的分子基础研究
基本信息
- 批准号:13440240
- 负责人:
- 金额:$ 10.75万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (B)
- 财政年份:2001
- 资助国家:日本
- 起止时间:2001 至 2002
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Autotrophic capacity of plants depends on a series of redox of redox metabolisms. An electron carrir protein, ferredoxin (Fd) and Fd-dependent enzymes function in combination as a molecular machinery for such metabolisms. The process of electron partitioning via Fd to various enzymes within plastids is crucial for photosynthetic activity and elucidation of its molecular mechanism is the main aim of this study. Following results has been obtained.1) X-ray crystsl structure of an electron transfer complex of root Ed and Fd:NADP+ reductase has been determied and this structure is found to be different from that of lesf counterpart.2) Electron transfer between sulfite reductase and Fd was investigate using wild type ans their mutant molecules and redox potential of Fd was found to be important for activity and substrate specificity of sulfite reductase. X-ray crystal analysis of this enzyme is now ongoing.3) Site diredted mutagenesis study of Fd-dependent glutamate synthase was done. Amino acids necessary for the activity of he amidotransferae domain of this enzyme were identified.4) Based on the total total genome information of Arabidopsis, 4 Fd isoprotiens were prepared and purified by reconbinant technique. Their abilities in the electron transfer to photosystem. I, Fd:NADP+ reductase and sulfite reductase were significantly different and physiological differentiation of their role was elucidated. Now proteins which interact with each Fd isoprotein are under investigation.5) A cyanobacterial mutant lacking Fd-dependent glutamate synthase exhibits unbalancing of carbon and nitrogen assimilation probably due to disturbance of partitioning of reducing equivalent. In response to this unbalancing the expression of glutamate synthase gene was up and down-regulated. We could monitor this gene expression by reporterassay syatem.
植物的自养能力取决于一系列氧化还原代谢的氧化还原过程。电子载体蛋白、铁还蛋白(Fd)和依赖于Fd的酶结合在一起,就像是这种代谢的分子机制。电子通过FD分配给叶绿体内各种酶的过程是光合作用活性的关键,阐明其分子机制是本研究的主要目的。研究结果如下:1)测定了根ED和FD的电子转移络合物的X射线晶体结构,发现该结构不同于LEF的结构。2)用野生型ANS研究了亚硫酸盐还原酶和FD之间的电子转移,发现FD的突变分子和氧化还原电位对亚硫酸盐还原酶的活性和底物专一性有重要影响。对该酶进行了X射线晶体分析。3)对依赖FD的谷氨酸合成酶进行了定点突变研究。4)根据拟南芥全基因组信息,利用重组技术制备并纯化了4个Fd同源蛋白。它们在电子转移到光系统中的能力。I、FD:NADP+还原酶和亚硫酸盐还原酶有显著差异,并阐明了它们的生理分化作用。5)缺乏依赖FD的谷氨酸合成酶的蓝藻突变体表现出碳氮同化失衡,这可能是由于还原当量的分配受到干扰所致。为了应对这种失衡,谷氨酸合成酶基因的表达受到了上调和下调。我们可以通过报告分析系统来监测该基因的表达。
项目成果
期刊论文数量(30)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
栗栖源嗣: "植物プラスチドのレドックス代謝の分子基盤"化学と生物. 39巻・10号. 660-666 (2001)
Genji Kurisu:“植物质体氧化还原代谢的分子基础”,化学与生物学,第 39 卷,第 10 期。660-666(2001 年)。
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Kada,S., Koike,H., Satoh,K., Hase, T., and Fujita,Y.: "Arrest of chlorophyll synthesis and differential decrease of photosystems I and II in a cyanobacterpal mutant lacking light-iudependent protochlorophyllide reductase"Plant Mot. Biol.. 51. 225-235 (200
Kada,S.、Koike,H.、Satoh,K.、Hase, T. 和 Fujita,Y.:“缺乏光依赖性原叶绿素还原酶的蓝细菌突变体中叶绿素合成的抑制和光系统 I 和 II 的差异性减少”
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長谷 俊治: "植物のパワージェネレーターは光でコントロールされる"第16回「大学と科学」公開シンポジウム組織委員会編『明日を拓く植物科学:光エネルギーを生物エネルギーにかえる植物の設計図を読む』. 69-77 (2002)
长谷俊二:《植物发电器受光控制》第十六届“大学与科学”公共研讨会组委会编辑《开拓明天的植物科学:解读将光能转化为生物能的植物蓝图》.69-77 (2002)
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栗栖 源嗣: "酵素との電子伝達複合体の構造解明"化学と生物. 39. 660-666 (2001)
Genji Kurisu:“用酶阐明电子传递复合物的结构”化学与生物学 39. 660-666 (2001)。
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Maeda,M., Hamada,D., Hoshino,M., Onda,Y., Hase,T., and Goto,Y.: "Partially folded structure of flavin adenine dinucleotide-depleted ferredoxin-NADP^+ reductase with residual NADP^+ binding domain"J. Biol. Chem.. 277. 17101-17107 (2002)
Maeda,M.、Hamada,D.、Hoshino,M.、Onda,Y.、Hase,T. 和 Goto,Y.:“黄素腺嘌呤二核苷酸耗尽的铁氧还蛋白-NADP^还原酶的部分折叠结构,具有残留的 NADP^
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HASE Toshiharu其他文献
HASE Toshiharu的其他文献
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{{ truncateString('HASE Toshiharu', 18)}}的其他基金
Analysis of a new substrate recognition mechanism of glutamine synthetase and its application for modulation of herbicide sensitivity
谷氨酰胺合成酶新底物识别机制分析及其在除草剂敏感性调节中的应用
- 批准号:
23651219 - 财政年份:2011
- 资助金额:
$ 10.75万 - 项目类别:
Grant-in-Aid for Challenging Exploratory Research
Regulation of plant redox metabolic function based on the atomic structure of proteins
基于蛋白质原子结构调控植物氧化还原代谢功能
- 批准号:
20370022 - 财政年份:2008
- 资助金额:
$ 10.75万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Molecular machinery regulating energy and metabolic networks in plant cells
调节植物细胞能量和代谢网络的分子机器
- 批准号:
15GS0320 - 财政年份:2003
- 资助金额:
$ 10.75万 - 项目类别:
Grant-in-Aid for Creative Scientific Research
Functional design of enzymes for glutamate synthesis and factors involved in cooperative assimilation of carbon and nitrogen
谷氨酸合成酶的功能设计及碳氮协同同化相关因素
- 批准号:
10640630 - 财政年份:1998
- 资助金额:
$ 10.75万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Analysis and modulation of photosynthetic metabolism through the mutagenesis of ferredoxin
通过铁氧还蛋白诱变分析和调节光合代谢
- 批准号:
08640828 - 财政年份:1996
- 资助金额:
$ 10.75万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Biochemical and molecular biological studies of electron partitioning among enzymes involved in the assimilation of inorganic compounds in plants
参与植物无机化合物同化的酶之间电子分配的生化和分子生物学研究
- 批准号:
05454019 - 财政年份:1993
- 资助金额:
$ 10.75万 - 项目类别:
Grant-in-Aid for General Scientific Research (B)
Expression of ferredoxin gene family and its physiological significance
铁氧还蛋白基因家族的表达及其生理意义
- 批准号:
02660086 - 财政年份:1990
- 资助金额:
$ 10.75万 - 项目类别:
Grant-in-Aid for General Scientific Research (C)
Molecular mechanisms of protein sorting between the mitochondrial outer and inner membranes
线粒体外膜和内膜之间蛋白质分选的分子机制
- 批准号:
63580151 - 财政年份:1988
- 资助金额:
$ 10.75万 - 项目类别:
Grant-in-Aid for General Scientific Research (C)
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线粒体功能障碍:了解功能失调的线粒体氧化还原代谢如何支撑病理学
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10272745 - 财政年份:2021
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Expanding the set of genetically encoded tools for compartment-specific manipulation of redox metabolism in living cells
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10437022 - 财政年份:2021
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Expanding the set of genetically encoded tools for compartment-specific manipulation of redox metabolism in living cells
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Defining and targeting the compartmentalization of redox metabolism in aging using novel genetically encoded tools
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Selective targeting of tumor cell redox metabolism and DNA damage responses to enhance cancer therapy
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