CAREER: Bioinorganic Redox Chemistry and Protein-Protein Interactions at an Electrode

智力上的优点。微生物，如欧洲亚硝化单胞菌和希瓦氏菌oneidensis能够显着的生化转化作为其整体代谢的一部分。像所有生物一样，微生物通过精心设计的方案利用能量，其中电子从一个蛋白质分子移动到另一个，形成一种内部电路。传统上，确定细胞内部电子工作的组成和电路是相当困难的。在本研究中，一种新的分析工具，蛋白质膜伏安法，将被用来了解的基本方式N。europaea和S. oneidensis在其内部电路的组件内移动电子。 具体而言，本项目将报告作为单个纯化蛋白质和大分子复合物的单血红素和多血红素细胞色素的氧化/还原（氧化还原）特性。将研究一系列多血红素电子转移蛋白（CymA，MtrA，STC，cyt c554和cyt cm 552），以了解氧化还原协同性在这些蛋白质的功能中发挥的潜在作用。 然后，将制备模型蛋白质-蛋白质复合物并进行电化学分析，以确定蛋白质-蛋白质相互作用如何影响和调节生物电子转移的热力学和动力学。最终，该项目将集中在确定蛋白质电化学如何可用作鉴定不稳定的氧化还原活性蛋白质-蛋白质复合物的快速工具。 更广泛的影响。 该项目将提供重要的见解功能，不稳定的蛋白质-蛋白质相互作用，以及帮助蛋白质电化学的发展作为一种分析工具。 了解N. europaea和S. oneidensis将使这些生物体在生物能源科学中得到利用。 此外，多学科研究将与教育计划一起进行，该计划将开发（1）一个新的第一年化学课程，提供跨学科，以问题和研究为导向的学习环境;（2）生物无机化学多学科研究生课程;（3）研究生研究行为和道德的系列研讨会，以及（4）本科和高中夏季研究的管道。

Intellectual merit. Microbes such as Nitrosomonas europaea and Shewanella oneidensis are capable of remarkable biochemical transformations as part of their overall metabolism. Like all living things, micro-organisms harness energy by elaborate schemes in which electrons move from one protein molecule to another, forming a kind of internal circuitry. Traditionally, the determination of the composition and circuitry of the inner electrical workings of a cell would be quite difficult. In this research, a new analytical tool, protein film voltammetry, will be used to learn about the fundamental ways N. europaea and S. oneidensis move electrons within the components of their internal circuitry. Specifically, this project will report upon the oxidation/reduction (redox) properties of monoheme and multiheme cytochromes as individual purified proteins and macromolecular complexes. A series of multiheme electron transfer proteins (CymA, MtrA, STC, cyt c554, and cyt cm552) will be studied to understand the potential role redox-cooperativity plays in the function of these proteins. Then, model protein-protein complexes will be prepared and analyzed electrochemically, to determine how protein-protein interactions influence and modulate the thermodynamics and kinetics of biological electron transfer. Ultimately, this project will focus on determination of how protein electrochemistry can be used as a rapid tool for the identification of labile, redox-active protein-protein complexes. Broader impacts. This project will provide important insights into functional, labile protein-protein interactions as well as aid the development of protein electrochemistry as an analytical tool. Understanding the complex electrochemical pathways of N. europaea and S. oneidensis will enable the exploitation of these organisms in the bioenergy sciences. Further, the multi-disciplinary research will proceed alongside an educational plan that will develop (1) a new, first-year chemistry course that provides an interdisciplinary, problem- and research-oriented learning environment; (2) multi-disciplinary graduate courses in bioinorganic chemistry; (3) a seminar series in graduate research conduct and ethics, and (4) a conduit for undergraduate and high-school summer research.