Studies on Factors for Controlling Electron Transfer Reactions and Biological Functions of Metalloproteins Using Functional Electrode Surfaces

利用功能电极表面研究电子转移反应的控制因素和金属蛋白的生物学功能

• 批准号: 11450329
• 负责人: TANIGUCHI Isao
• 金额: $ 9.34万
• 依托单位: Kumamoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11450329/
• 关键词: Metalloprotein; Bio-functional electrode; Single crystal electrode; Surface structure; Electron transfer reaction; Myoglobin; Ferredoxin; Mutated molecule

In the present study, using surface modified single crystal gold electrodes and other functionalized electrodes, electron transfer reactions and biological functions of metalloproteins have been examined.1. Using thermo-stable Chlorella ferredoxin as an electron transfer mediator of enzyme, artificial photosynthetic electrodes, on which enzyme and ferredoxin were co-immobilized in poly-peptide thin films, were prepared.2. Functional electrode surfaces modified with SAMs of 2-, 3-, and 4-pyridinethiols and 2,2' and 5,5' pyrimidinethiols were examined at molecular level to understand their structures and functions for electron transfer reactions of metalloproteins by STM, surface-enhanced IR absorption spectroscopy (SEIRAS), surface-enhanced Raman spectroscopy (SERS), impedance measurement, contact angle measurement, the electrochemical reductive desorption technique, etc. On the Au(111) surface, for example, adsorbed 4 pyridinethiol (4-PySH) showed the rectangular unit cell of p(5 x 【sq … More uare root】3R-30°) structure with an interaction of two neighboring 4-PySH, while the 2-PySH modified electrode showed a p(4 x 【square root】7R-40.9°) structure with no dimer formation. 3-PySH showed a p(6 x 【square root】3R-30°) structure. Similar structures were also observed on the Au(100) surface. The observed electrochemical response of cytochrome c can be explained in terms of the observed structures of modified electrodes. However, properties of the surfaces required for the rapid electron transfer depended on metalloproteins used.3. Using highly hydrophilic surface of In2O3 electrodes, biological functions and electron transfer kinetics of native and reconstituted myoglobins were examined in light and heavy water. The rates of electron transfer reactions of myoglobins were related to the porphyrin backbone of the heme, type of metal ion of the redox center, coordination structure etc. Biological functions depended on heme structures. New reconstituted myoglobin molecules having biological functions better than native myoglobin were also prepared.4. A new artificial heme-incorporated polypeptide was prepared, and modified on the electrode surface to give electron-transfer reactions similar to a metalloprotein. Less

本研究利用表面修饰的单晶金电极和其他功能化电极，对金属蛋白的电子转移反应和生物学功能进行了研究。 1．以热稳定性小球藻铁氧化还原蛋白作为酶的电子传递介体，制备了酶与铁氧化还原蛋白共固定在多肽薄膜上的人工光合电极。 2．通过 STM、表面增强红外吸收光谱 (SEIRAS)、表面增强拉曼光谱 (SERS)、阻抗，在分子水平上检查用 2-、3-和 4-吡啶硫醇以及 2,2' 和 5,5' 嘧啶硫醇 SAM 修饰的功能电极表面，以了解它们在金属蛋白电子转移反应中的结构和功能 以Au(111)表面为例，吸附的4吡啶硫醇(4-PySH)呈现出p(5 x 【sq … More uare root】3R-30°)结构的矩形晶胞，并与两个相邻的4-PySH相互作用，而2-PySH修饰电极则呈现出p(4 x 【平方根】7R-40.9°）结构，无二聚体形成。 3-PySH呈p(6 x 【平方根】3R-30°)结构。在 Au(100) 表面也观察到类似的结构。观察到的细胞色素 c 的电化学响应可以用观察到的修饰电极的结构来解释。然而，快速电子转移所需的表面特性取决于所使用的金属蛋白。3．利用 In2O3 电极的高亲水表面，在轻水和重水中检查了天然和重构肌红蛋白的生物功能和电子转移动力学。肌红蛋白的电子转移反应速率与血红素的卟啉骨架、氧化还原中心的金属离子类型、配位结构等有关。生物功能取决于血红素的结构。还制备了新的重构肌红蛋白分子，其生物学功能优于天然肌红蛋白。 4．制备了一种新的人工血红素掺入多肽，并在电极表面进行修饰，以产生类似于金属蛋白的电子转移反应。较少的

项目成果

S. Uemura, A. Ohira, T. Ishizaki, M. Sakata, M. Kunitake, I. Taniguchi, C. Hirayama: "In situ STM Visualization of Fullerlene Epitaxial Adlayers on Au(111) Surfaces Prepared by the Transfer of Langmuir Films"Chem. Lett.. 279-280 (1999)

S. Uemura、A. Ohira、T. Ishizaki、M. Sakata、M. Kunitake、I. Taniguchi、C. Hirayyama：“通过 Langmuir 薄膜转移制备的 Au(111) 表面富勒烯外延粘附层的原位 STM 可视化

谷口,功: "ポイントミューテーションによるタンパク質の電子機能の改変"電気化学. 68(5). 341-347 (2000)

Taniguchi, Isao：“点突变对蛋白质电子功能的修饰”，电化学 68(5)。

Taniguchi,Isao: "Bioelectroanalytical Chemistry Using Functionalized Electrodes : Fundamentals and Applicqations"Proc.Workshop cum Seminar on Electroanaltytical Chemistry and Allid Topics(ELAC-2000). 1. 118-23 (2000)

Taniguchi,Isao：“使用功能化电极的生物电分析化学：基础知识和应用”Proc.研讨会暨电分析化学和所有主题研讨会（ELAC-2000）。

Uemura,Shinobu: "Novel "wet process" technique based on electrochemical replacement for the preparation of fullerene epitaxial adlayers"Langmuir. 17. 5-7 (2001)

Uemura，Shinobu：“基于电化学替代的新型“湿法”技术用于制备富勒烯外延吸附层”Langmuir。

Yoshimoto,Soichiro: "Direct Observation of Structural Change Induced by Redox Reaction of Bis(2-Anthraquinyl)disulfide SAM on Au(100)-(1x1) by in situ High Resoltion STM"Langmuir,. (in press). (2000)

Yoshimoto，Soichiro：“通过原位高分辨率 STM 直接观察 Au(100)-(1x1) 上双（2-蒽醌基）二硫化物 SAM 的氧化还原反应引起的结构变化”Langmuir，。