Elucidation of molecular mechanism of bilin synthesis by ferredoxin-dependent bilin reductases
阐明铁氧还蛋白依赖性胆素还原酶合成胆素的分子机制
基本信息
- 批准号:20370037
- 负责人:
- 金额:$ 12.81万
- 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:2008
- 资助国家:日本
- 起止时间:2008 至 2010
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
In order to elucidate the molecular mechanism of PcyA, we determined the crystal structures of PcyA in complex with the reaction intermediate (18^1,18^2-dihydrobiliverdin), PcyA in complex with the synthetic analog of substrate (biliverdin XIIIα), and PcyA mutant protein of E76Q in complex with BV. These structural studies showed that the carboxyl group of Glu76 in PcyA-BV was in close proximity to the D-ring vinyl group, suggesting the presence of OH...π hydrogen bond here and that this bond is a key for the preceding reduction of the D-ring vinyl group.When Val225 was replaced by Asp, the absorption spectrum of this mutant protein in complex with BV was very different from that of PcyA-BV. The orientation of BV in V225D-BV was inverted and the induced-fit did not occur upon BV binding. This observation presented a cautionary note about interpreting functional data derived from a mutated protein in the absence of its exact structure. In parallel we expressed, purified and crystallized cyanobacterial biliverdin reductase, an enzyme that reduces at different site of BV from PcyA, We also prepared and crystallized the Se-Met form of biliverdin reductase, and performed preliminary X-ray diffraction analyses for these crystals.We determined the crystal structure of red chlorophyll catabolite reductase (RCCR), a key enzyme in the chlorophyll breakdown pathway. The structure was essentially the same as PcyA, implicating that RCCR belongs to the ferredoxin-dependent bilin reductase family. We also determined the crystal structures of RCCR and its mutant protein in complex with substrate, proposing the reaction mechanism as well as the active residues of RCCR.
为了阐明PcyA的分子机制,我们测定了PcyA与反应中间体(18^1,18^2-二氢胆绿素)复合物、PcyA与合成底物类似物(胆绿素XIIIα)复合物以及PcyA突变体蛋白E76 Q与BV复合物的晶体结构。这些结构研究表明PcyA-BV中Glu 76的羧基与D-环乙烯基非常接近,表明存在OH。当Val 225被Asp取代后,该突变蛋白与BV复合物的吸收光谱与PcyA-BV的吸收光谱有很大不同。BV在V225 D-BV中的取向是颠倒的,并且在BV结合时不发生诱导拟合。这一观察结果提出了一个警告,说明在缺乏确切结构的情况下解释来自突变蛋白质的功能数据。同时,我们表达、纯化并结晶了蓝藻胆绿素还原酶(一种在BV的不同位点还原PcyA的酶),制备并结晶了胆绿素还原酶的Se-Met形式,并对这些晶体进行了初步的X射线衍射分析,确定了叶绿素分解途径中的关键酶红色叶绿素分解代谢物还原酶(RCCR)的晶体结构。其结构与PcyA基本相同,表明RCCR属于铁氧还蛋白依赖性胆色素还原酶家族。我们还测定了RCCR及其突变蛋白与底物复合物的晶体结构,提出了RCCR的反应机理和活性残基。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Crystal structure of rat heme oxygenase-1 in complex with ferrous verdoheme : presence of a hydrogen bond network on the distal side
大鼠血红素加氧酶-1与亚铁绿血红素复合物的晶体结构:远端存在氢键网络
- DOI:
- 发表时间:2009
- 期刊:
- 影响因子:0
- 作者:Sato;H.;Sugishima;M.;Sakamoto;H.;Higashimoto;Y.;Shimokawa;C.;Fukuyama;K.;and Noguchi;M.
- 通讯作者:M.
グルタチオン代謝の鍵酵素γ-グルタミルトランスペプチダーゼに対する古典的阻害剤アシビシンの結合様式
经典抑制剂阿西维星与谷胱甘肽代谢关键酶γ-谷氨酰转肽酶的结合模式
- DOI:
- 发表时间:2010
- 期刊:
- 影响因子:0
- 作者:伊田知代;和田啓;平竹潤;鈴木秀之;福山恵一
- 通讯作者:福山恵一
Crystal structure of the halotolerant γ-glutamyltranspeptidase from Bacillus subtilis in complex with glutamate reveals its unique architecture of the solvent-exposed catalytic pocket
来自枯草芽孢杆菌的耐盐γ-谷氨酰转肽酶与谷氨酸复合物的晶体结构揭示了其溶剂暴露催化袋的独特结构
- DOI:
- 发表时间:2010
- 期刊:
- 影响因子:0
- 作者:K.Wada;M.Irie;H.Suzuki;K.Fukuyama
- 通讯作者:K.Fukuyama
鉄硫黄クラスター生合成系における硫黄の供給メカニズム解明に向けた構造生物学的アプローチ
阐明铁硫簇生物合成系统中硫供应机制的结构生物学方法
- DOI:
- 发表时间:2010
- 期刊:
- 影响因子:0
- 作者:平林佳;中嘉孝;高橋康弘;福山恵一;和田啓
- 通讯作者:和田啓
Enhancement of glutaryl-7-aminocepharosporanic acid acylase activity of γ-glutamyltranspeptidase of Bacillus subtilis
枯草芽孢杆菌γ-谷氨酰转肽酶戊二酰-7-氨基头孢酸酰基转移酶活性的增强
- DOI:
- 发表时间:2010
- 期刊:
- 影响因子:4.7
- 作者:H.Suzuki;K.Fukuyama;et al.
- 通讯作者:et al.
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FUKUYAMA Keiichi其他文献
FUKUYAMA Keiichi的其他文献
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{{ truncateString('FUKUYAMA Keiichi', 18)}}的其他基金
Elucidation of mechanisms of bilin synthesizing enzyme and photo-adaptation regulating protein
胆素合成酶和光适应调节蛋白机制的阐明
- 批准号:
23370052 - 财政年份:2011
- 资助金额:
$ 12.81万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Elucidation of reaction mechanism ofthe enzymes involved in the syntbssis of photosynthetic pigments
光合色素合成酶反应机制的阐明
- 批准号:
18570105 - 财政年份:2006
- 资助金额:
$ 12.81万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Functional analysis of proteins based on the tertiary structures focussing on hydrogen atom positions
基于以氢原子位置为重点的三级结构的蛋白质功能分析
- 批准号:
14580674 - 财政年份:2002
- 资助金额:
$ 12.81万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Mechanism of DNA excision repair based on the three-dimensional structure of UvrABC endonuclease
基于UvrABC核酸内切酶三维结构的DNA切除修复机制
- 批准号:
12680659 - 财政年份:2000
- 资助金额:
$ 12.81万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Analysis of assembly mechanism of spherical virus based on its high resolution three-dimensional structure
基于高分辨率三维结构的球形病毒组装机制分析
- 批准号:
10680634 - 财政年份:1998
- 资助金额:
$ 12.81万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Crystallographic and Genetic Study of Cytochrome bc1 Complex
细胞色素bc1复合物的晶体学和遗传学研究
- 批准号:
08044203 - 财政年份:1996
- 资助金额:
$ 12.81万 - 项目类别:
Grant-in-Aid for international Scientific Research
Study on the reaction mechanism of peroxidase based on its tertiary structure
基于过氧化物酶三级结构的反应机理研究
- 批准号:
06680654 - 财政年份:1994
- 资助金额:
$ 12.81万 - 项目类别:
Grant-in-Aid for General Scientific Research (C)
Studies on the reaction mechanism of peroxidase based on its tertiary structure
基于过氧化物酶三级结构的反应机理研究
- 批准号:
04680055 - 财政年份:1992
- 资助金额:
$ 12.81万 - 项目类别:
Grant-in-Aid for General Scientific Research (C)
X-Ray Crystallographic Analysis of Spherical Plant Viruses
球形植物病毒的 X 射线晶体学分析
- 批准号:
61580049 - 财政年份:1986
- 资助金额:
$ 12.81万 - 项目类别:
Grant-in-Aid for General Scientific Research (C)
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- 批准号:
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