Study on Bioloigical Functions Related to Dynamic Structural Change of Metalloproteins Using Newly Developed Spectroelectrochemical Techniques

利用新开发的光谱电化学技术研究与金属蛋白动态结构变化相关的生物功能

基本信息

批准号：
04453082
负责人：
TANIGUCHI Isao
金额：
$ 4.61万
依托单位：
Kumamoto Uiversity
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1992
资助国家：
日本
起止时间：
1992 至 1994
项目状态：
已结题

项目摘要

The aims of the present study are to i) prepare of optically transparent functional electrodes for Spectroelectrochemical measurements, ii) develop of new spectroelectrochemical techniques, and iii) on the basis of these developments, study on biological functions related to structural change of metalloproteins.Preparation of functional electrodes for metalloprotein electrochemistryWe have succeeded to prepare optically transparent In_2O_3 functionalized electrodes for myoglobins and ferredoxins. The surface hydrophilicity of an In_2O_3 electrode affected very much the perk separation of the voltammogram of myoglobin, The water layr of the electrode surface may prevent from the strong adsorption of myoglobin to exclude denaturation of the protein, and also surface water would preferably take part in the redox reaction (water molecule acts as the sixth ligand of metmyoglobin). On the other hand, at positively charged polypeptide modified electrodes, for example, negatively charged ferre … More doxins showed clear voltammograms. Such surface functions as charge, hydrophilicity and strength of interaction between protein and electrode, are found to be important, but importantly the required degrees of these functions depend on the proteins and functional electrode surfaces of interest.Development of new spectroelectrochemical techniquesUsing functionalized optically transparent electrodes, in situ UV-visible, Near Infratred (NIR), circular dichroism (CD) and magnetic circular dichroism (MCD) spectroelectrochemical cells were prepared. Also, by modification of the instruments, data accumulation and automatic measurements became possible to obtain reliable and reproducible data with high S/N ratios.Spectroelectrochemical measurements of metallopreteinsUsing the developed techniques in the present study, redox reactions of cytochrome c, myoglobins, ferredoxins and their mutated molecules were examined to study in situ the dynamics of structural change during electron transfer related to biological functions of these metalloprtoteins and to monitor the bioelectrochemical reactions such as NADPH production in the presence of ferredoxin-NADP^+-reductase (FNR) using thermostable chlorella ferredoxin as a mediator. Less

本研究的目的是i）准备光透明的功能电极以进行脑光谱测量，ii）根据这些发展，开发了新的Spectroelectroectrogation技术，以及III）。 IN_2O_3肌球蛋白和铁蛋白的功能化电极。 IN_2O_3电极的表面亲水性非常影响肌红蛋白的伏安图的分离，电极表面的水层可能会阻止肌球蛋白的强添加蛋白质，以排除蛋白质的变性，并且表面水也会参与氧化还原反应（水中六叶）的反应（sixy ligigy）。另一方面，例如，在带正电的多肽修饰的电子中，带负电荷的Ferre…更多的毒素显示清晰的伏安图。 Such surface functions as charge, hydrophilicity and strength of interaction between protein and electrode, are found to be important, but importantly the required degrees of these functions depend on the proteins and functional electrode surfaces of interest.Development of new spectroelectrochemical techniquesUsing functionalized optically transparent electrodes, in situ UV-visible, Near Infratred (NIR), circular dichroism (CD)制备了磁圆二色性（MCD）光谱细胞。同样，通过修改仪器，数据积累和自动测量是可能的，以获得具有高S/N比的可靠且可重复的数据。在本研究中，在本研究中对金属剂的光谱测量进行了测量，在本研究中开发的技术，细胞染料C，肌球蛋白，纤维素，纤维素氧化物的构建过程的氧化还原反应，对肌球蛋白，纤维素的变化进行了研究，这些反应是对肌球蛋白，纤维素的构成尺寸，效应了分类的分子尺寸，以下过程转移与这些金糖蛋白的生物学功能相关，并使用热蛋白-NADP^+ - 还原酶（FNR）在存在下使用热蛋白溶质性小氯甲烷铁氧还蛋白作为介体来监测诸如NADPH产生的生物化学反应。较少的