Analysis of an enzymatic domain that is involved in specificity of phosphoryl donors, polyphosphate and ATP
参与磷酰基供体、多磷酸盐和 ATP 特异性的酶结构域分析
基本信息
- 批准号:15560676
- 负责人:
- 金额:$ 2.37万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (C)
- 财政年份:2003
- 资助国家:日本
- 起止时间:2003 至 2004
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Inorganic polyphosphate (polyP) is a linear polymer of phosphate linked by high energy phosphoanhydride bonds. PolyP is found in abundance in volcanic condensates. Therefore, some investigators consider that polyP may be energy currency for prebiotic life. We found that a strictly polyP-dependent glucokinase in Microlunatus phosphovorus. ATP is used to phosphorylate glucose in many organisms, while polyP is used in this microorganism. We determined a primary structure of this polyP-glucokinase, and found that the active site residues are conserved in both of ATP- and polyP-glucokinase. However, a domain IIB is only observed in ATP-glucokinase. We considered that ATP-glucokinase is evolved from polyP-glucokinase. In this study, we wanted to reveal structural changes in glucokinase when it acquire the domain IIB and changes its phosphoryl donor. First we purified polyP-glucokinase from E.coli recombinant. The purified enzyme is further concentrated and used in order to find crystalization condition. Finally we succeeded in crystalization of polyP-glucokinase and obtaining X-ray diffraction pattern with two-angstrom resolution. We have not yet solved, but succeeded in predicting its three-dimensional structure. We predicted that polyP-glucokinase acquired adenosine recognizing domain and evolved to ATP-glucokinase.
无机聚磷酸盐(polyP)是由高能磷酸酐键连接的磷酸盐的线性聚合物。PolyP在火山冷凝物中含量丰富。因此,一些研究人员认为聚磷可能是前生物生命的能量货币。我们发现,在Microlunatus phosphovorus严格聚P依赖的葡萄糖激酶。在许多生物中,ATP用于磷酸化葡萄糖,而聚P用于这种微生物。我们确定了这种聚P-葡萄糖激酶的一级结构,并发现活性位点残基在ATP-和聚P-葡萄糖激酶中是保守的。然而,结构域IIB仅在ATP-葡萄糖激酶中观察到。我们认为ATP-葡萄糖激酶是由聚P-葡萄糖激酶进化而来的。在这项研究中,我们希望揭示葡萄糖激酶在获得结构域IIB并改变其磷酰基供体时的结构变化。首先,我们从大肠杆菌重组体中纯化了聚P-葡萄糖激酶。将纯化的酶进一步浓缩并用于寻找结晶条件。最后,我们成功地结晶了聚P-葡萄糖激酶,并获得了两埃分辨率的X射线衍射图。我们尚未解决,但成功预测了它的三维结构。我们预测聚P-葡萄糖激酶获得腺苷识别结构域并进化为ATP-葡萄糖激酶。
项目成果
期刊论文数量(7)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
黒田章夫: "微生物におけるポリリン酸代謝制御機構の解明と利用"生物工学会誌. 81. 104-111 (2003)
Akio Kuroda:“微生物中多磷酸盐代谢控制机制的阐明和利用”日本生物工程学会杂志 81. 104-111 (2003)。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
New Aspects of Polyphosphate Functions and Applications in Microorganisms
聚磷酸盐功能的新方面及其在微生物中的应用
- DOI:
- 发表时间:2004
- 期刊:
- 影响因子:0
- 作者:Akio Kuroda
- 通讯作者:Akio Kuroda
Tanaka S. et al.: "Strictly polyphosphate-dependent glucokinase in a polyphosphate-accumulating bacterium Microlunatus phosphovorus"Journal of Bacteriology. 185. 5654-5656 (2003)
Tanaka S.等人:“聚磷酸盐积累细菌Microlunatus phosphorus中的严格聚磷酸盐依赖性葡萄糖激酶”细菌学杂志。
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- 发表时间:
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- 影响因子:0
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KURODA Akio其他文献
KURODA Akio的其他文献
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{{ truncateString('KURODA Akio', 18)}}的其他基金
Novel method utilizing bisulfite conversion with dual amplification-refractory mutation system polymerase chain reaction to detect circulating pancreatic beta cell cfDNA.
利用亚硫酸氢盐转化和双重扩增-难治性突变系统聚合酶链反应检测循环胰腺 β 细胞 cfDNA 的新方法。
- 批准号:
18K07448 - 财政年份:2018
- 资助金额:
$ 2.37万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Detection of pancreatic beta cell DNA in the circulation utilizing epigenetic modification
利用表观遗传修饰检测循环中的胰腺 β 细胞 DNA
- 批准号:
15K06910 - 财政年份:2015
- 资助金额:
$ 2.37万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Application of a transgenic plant conferred with phosphite-oxidizing ability to efficient utilization of phosphite waste
具有亚磷酸盐氧化能力的转基因植物在亚磷酸盐废物高效利用中的应用
- 批准号:
25630382 - 财政年份:2013
- 资助金额:
$ 2.37万 - 项目类别:
Grant-in-Aid for Challenging Exploratory Research
ATP amplification reaction in pico-liter chamber and its application to bacteria detection.
皮升室中ATP扩增反应及其在细菌检测中的应用
- 批准号:
23656529 - 财政年份:2011
- 资助金额:
$ 2.37万 - 项目类别:
Grant-in-Aid for Challenging Exploratory Research
Development of NADH regeneration system by thermostable phosphite dehydrogenase
耐热亚磷酸脱氢酶NADH再生系统的开发
- 批准号:
22360346 - 财政年份:2010
- 资助金额:
$ 2.37万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Development of an ultra-sensitive ATP assay by an enzymatic ATP amplification reaction.
通过酶促 ATP 扩增反应开发超灵敏 ATP 测定。
- 批准号:
13650858 - 财政年份:2001
- 资助金额:
$ 2.37万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Study on microbial proteiolipid involved in haydroxyapatite formation.
微生物蛋白脂参与羟基磷灰石形成的研究。
- 批准号:
11650819 - 财政年份:1999
- 资助金额:
$ 2.37万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
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