Structure and function of the heme-based gas sensor proteins

基于血红素的气体传感器蛋白的结构和功能

基本信息

  • 批准号:
    14380303
  • 负责人:
  • 金额:
    $ 9.73万
  • 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:
    2002
  • 资助国家:
    日本
  • 起止时间:
    2002 至 2003
  • 项目状态:
    已结题

项目摘要

HemAT-Bs is a heme-containing signal transducer protein responsible for aerotaxis of Bacillus subtilis. The recombinant HemAT-Bs expressed in Escherichia coli was purified as the oxy form in which oxygen was bound to the ferrous heme. Oxygen binding and dissociation rate constants were determined to be kon = 32 μM^<-1> s^<-1> and koff = 23^<-1>, respectively, revealing that HemAT-Bs has a moderate oxygen affinity similar to that of sperm whale myoglobin (Mb). The rate constant for autoxidation at 37 ℃ was 0.06 h^<-1>, which is also close to that of Mb. Although the electronic absorption spectra of HemAT-Bs were similar to those of Mb, HemAT-Bs showed some unique characteristics in its resonance Raman spectra. Oxygen-bound HemAT-Bs gave the Fe-0_2 band at a noticeably low frequency (560 cm^<-1>), which suggests a unique hydrogen bonding between a distal amino acid residue and the proximal atom of the bound oxygen molecule. Deoxy HemAT-Bs gave the Fe-His band at a higher frequency (225 cm<-1>) than those of ordinary His-coordinated deoxy heme proteins. CO-bound HemAT-Bs gave the Fe-CO and C-O bands at 494 and 1964 cm1, respectively, which fall on the same C-O versus Fe-CO correlation line as that of Mb. Based on these results, the structural and functional properties of HemAT-Bs has been elucidated.In this work, a new CooA homologue from C.hydrogenoformans (Ch-CooA) was also studied. Ch-CooA also contains a protoheme as the active site for sensing CO. The heme in Ch-CooA is six-coordinate, low spin state in the ferric, ferrous and CO-bound forms. The heme environmental structure of Ch-CooA was elucidated by mutagenesis and spectroscopic studies.
HemAT-Bs是一种含血红素的信号转导蛋白,负责枯草芽孢杆菌的嗜氧性。在大肠杆菌中表达的重组HemAT-Bs被纯化为氧结合亚铁血红素的氧合形式。测得的氧结合和解离速率常数分别为kon = 32 μM^<-1>s^<-1>和koff = 23^<-1>,表明HemAT-Bs具有与抹香鲸肌红蛋白(Mb)相似的中等氧亲和力。在37 ℃下的自氧化速率常数为0.06 h ~(-1)<-1>,也接近Mb。虽然HemAT-Bs的电子吸收光谱与Mb相似,但其共振拉曼光谱显示出一些独特的特征。氧结合的HemAT-Bs在明显低的频率(560 cm-2)处给出Fe-0_2带<-1>,这表明在结合的氧分子的远端氨基酸残基和近端原子之间存在独特的氢键。脱氧HemAT-Bs给出了铁-组氨酸带在更高的频率(225厘米<-1>)比普通的组氨酸协调脱氧血红素蛋白。CO结合的HemAT-Bs在494和1964 cm-1处分别给出了Fe-CO和C-O带,它们与Mb落在相同的C-O与Fe-CO相关线上。在此基础上,对HemAT-Bs的结构和功能特性进行了初步的研究,并对产氢梭菌的一个新的CooA同系物(Ch-CooA)进行了研究。Ch-CooA还含有一个血红素作为传感CO的活性位点。Ch-CooA中的血红素是六配位的低自旋态,以三价铁、亚铁和CO结合的形式存在。通过诱变和光谱研究阐明了Ch-CooA的血红素环境结构。

项目成果

期刊论文数量(50)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Shigetoshi Aono: "Resonance Raman and ligand binding analysis of the oxygen-sensing, signal transducer protein HemAT from Bacillus subtilis"Methods in Enzymol.. 381. 618-628 (2004)
Shigetoshi Aono:“来自枯草芽孢杆菌的氧传感信号转导蛋白 HemAT 的共振拉曼和配体结合分析”Enzymol.. 381. 618-628 (2004) 中的方法
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Shigetoshi Aono: "Resonance Raman and Ligand Binding Analysis of the Oxygen-sensing Signal Transducer Protein HemAT from Bacillus subtilis"Methods in Enzymology. 印刷中. (2004)
Shigetoshi Aono:“来自枯草芽孢杆菌的氧传感信号转导蛋白 HemAT 的共振拉曼和配体结合分析”酶学方法(2004 年)。
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Shigetoshi Aono: "Biochemical and Biophysical Properties of the CO-sensing transcriptional Activator CooA"Acc.Chem.Res.. 36・11. 825-831 (2003)
Shigetoshi Aono:“CO 感应转录激活剂 CooA 的生物化学和生物物理特性”Acc.Chem.Res. 36・11(2003)。
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    0
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Shigetoshi Aono: "Resonance Raman and ligand binding studies of the oxygen-sensing signal transducer protein Hem AT from Bacillus subtilis"J.Biol.Chem.. 277. 13528-13538 (2002)
Shigetoshi Aono:“来自枯草芽孢杆菌的氧传感信号转导蛋白 Hem AT 的共振拉曼和配体结合研究”J.Biol.Chem.. 277. 13528-13538 (2002)
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    0
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青野重利: "一酸化炭素センサーとして機能する転写調節因子CooAの構造と機能"Molecular Medicine. 40・2. 158-164 (2003)
Shigetoshi Aono:“作为一氧化碳传感器发挥作用的转录调节器 CooA 的结构和功能” 40・2(2003)。
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AONO Shigetoshi其他文献

AONO Shigetoshi的其他文献

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{{ truncateString('AONO Shigetoshi', 18)}}的其他基金

Construction of sensing systems for environmental pollutions based on biological sensing function
基于生物传感功能的环境污染传感系统构建
  • 批准号:
    25550047
  • 财政年份:
    2013
  • 资助金额:
    $ 9.73万
  • 项目类别:
    Grant-in-Aid for Challenging Exploratory Research
Construction of the homing cells harboring sensitive fluorescence probes for detection of environmental pollutants
构建含有敏感荧光探针的归巢细胞,用于检测环境污染物
  • 批准号:
    23651086
  • 财政年份:
    2011
  • 资助金额:
    $ 9.73万
  • 项目类别:
    Grant-in-Aid for Challenging Exploratory Research
Structure and function of sensor proteins that regulate biological functions by gas molecules
通过气体分子调节生物功能的传感器蛋白的结构和功能
  • 批准号:
    23350084
  • 财政年份:
    2011
  • 资助金额:
    $ 9.73万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Structure and function of metal-containing sensor proteins that use gas molecules as physiological effectors
使用气体分子作为生理效应器的含金属传感器蛋白的结构和功能
  • 批准号:
    19370059
  • 财政年份:
    2007
  • 资助金额:
    $ 9.73万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
The study on the structure and function of the transcriptionalregulator containing a heme as a signal sensor
含血红素信号传感器的转录调节因子的结构与功能研究
  • 批准号:
    12680631
  • 财政年份:
    2000
  • 资助金额:
    $ 9.73万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
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