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

知识价值。微生物如欧洲亚硝基单胞菌和希瓦氏菌能够显著的生化转化作为其整体代谢的一部分。像所有的生物一样，微生物通过复杂的计划来利用能量，电子从一个蛋白质分子移动到另一个蛋白质分子，形成一种内部电路。传统上，确定电池内部电工作的组成和电路是相当困难的。在这项研究中，一种新的分析工具，蛋白质膜伏安法，将被用来了解欧洲蕨和美国蕨在其内部电路组件内移动电子的基本方式。具体来说，本项目将报告单血红素和多血红素细胞色素作为单个纯化蛋白质和大分子复合物的氧化/还原（氧化还原）特性。我们将研究一系列多血红素电子转移蛋白（CymA、MtrA、STC、cyt c554和cyt cm552），以了解氧化还原-协同性在这些蛋白的功能中所起的潜在作用。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，该项目将专注于确定如何将蛋白质电化学用作鉴定不稳定，氧化还原活性的蛋白质-蛋白质复合物的快速工具。更广泛的影响。该项目将提供对功能，不稳定的蛋白质-蛋白质相互作用的重要见解，并有助于蛋白质电化学作为分析工具的发展。了解欧苔草和欧苔草复杂的电化学途径将有助于这些生物在生物能源科学中的开发利用。此外，多学科研究将与一项教育计划一起进行，该计划将开发：(1)新的一年级化学课程，提供跨学科、问题和研究导向的学习环境；(2)生物无机化学多学科研究生课程；(3)研究生研究行为和伦理系列研讨会；(4)本科生和高中暑期研究的渠道。

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.