蛋白质/酶中高氧化铁的研究是现代科学的热点课题之一。高氧化铁的生成和表征对于理解和认识蛋白质/酶的反应本质以及相应的生物氧化过程具有十分重要的意义。迄今为止，蛋白质中高氧化铁的产生都是利用底物或底物的类似物来实现的，反应是不可逆的，而通常产生的高氧化铁物种是暂态的或者亚稳态的，在分析过程中要特殊的捕获技术来满足分析要求。申请者结合自身多年的蛋白质电化学和光谱电化学研究背景，针对目前用于蛋白质/酶中的高氧化铁的研究方法的不足，提出采用基于介体型的现场光谱电化学来研究Heme蛋白质/酶的研究策略。并拟计划开展系列高氧化铁相关的电化学,光谱动力学研究。申请人有良好的工作基础，并做了必要的预研，本项目的研究方案可行性高。目前国际上还没有关于用电化学手段产生蛋白质/酶中高氧化铁的报道，本项目为蛋白质中高氧化铁的研究提供了一个新的思路，为了解高氧化铁相关的蛋白质/酶的生物学和化学本质提供了新的策略。

Investigation on highly oxidized iron species in proteins/enzymes currently is a promising reaserch topic. Generation and characterization of high-valent iron species are vital for understanding of the nature of such enzymatic reactions and corresponding biological oxidation process. So far, various methods have been extensively employed to investigate highly oxidized iron species intermediates in proteins. Among these methods, highly oxidized iron species were generated only in the reaction with substrates or their analogs, and thus the reaction is irreversible. Besides, many reactions make such species transient and metastable, which limits their population and requires trapping techniques. Compared with other methods, electrochemical methods are advantageous with respect to their rapid response time and simple procedures which have been proved to be a promising tool to understand the thermodynamics and kinetics of the electron transfer process, catalytic activities, and the relationship between structure and function of redox proteins. Moreover, electrochemistry method can be coupled relative easily to other techniques, such as spectroscopic techinque to constitute more powerful method, spectroelectrochemistry. In this project, we focus the the electrochemical generation and spectroelectrochemical characterization of high-valent iron speices in Heme protein based on using appropriate electron mediator to facilitate electron transfers between electrode and heme centers in proteins and thus conduct the ferric/ferryl electrochemistry. The utilization of electrochemical method to generate highly oxidized iron species has not been reported yet. This project is combined the protein electrochemistry and in-situ spectroelectrochemistry technology, which is expected to provide the electrochemical parameters (eg. Midpoint potential), kinetics parameters, and electron transfer pathway of protein during electrochemical redox process. The applicant has paved sound background knowledge and techniques for this project. Some preliminary investigations have been implemented, which indicated that the proposed approaches in this project are feasible. This study would definitely provide a novel strategy for for investigation of high-valent iron species in proteins which could potentially pave a facile approach to study the biology and biochemistry of protein associated with high-valent iron species.