CAREER: Towards the Rational Control of Redox Potential and Catalytic Activity of Designed Functional Membrane Proteins

在化学系有机动力学项目的支持下，亚利桑那州立大学的Giovanna Ghirlanda教授将设计和研究具有可调特性的新型功能膜蛋白。作为在这个方向上的第一步，血红素结合位点已被工程化到一个天然的膜蛋白，血型糖蛋白A，它形成稳定的二聚体，其跨膜螺旋结构域的协会。最初的设计，ME1，结合铁原卟啉IX具有高亲和力，表现出氧化还原电位的范围内的天然膜血红素结合蛋白，并显示出新生的催化活性。在资助期间，该系统将被进一步研究，并作为模板，用于设计一个大型突变体库，明确设计用于优化血红素结合和控制氧化还原电位。该文库将用于分析血红素环境的重要性，其疏水性和芳香族残基的存在，以确定ME1支架背景下的中点电位和结合亲和力。在这些结果的基础上，ME1将演变成一种人工酶的单加氧酶和过氧化物酶reactivity.The有机和大分子化学计划支持教授亚利桑那州州立大学的Giovanna Ghirlanda，其研究膜相关蛋白质是一个相对未开发的资源，作为生物传感器和生物催化剂的应用。对膜环境中蛋白质结构和功能之间关系的更深入理解将允许制备针对特定生物和非生物应用的人工小型化蛋白质。Ghirlanda博士将与亚利桑那州立大学的一个现有项目合作，实施一种综合方法，将K-12学校的课堂演示、本科生培训和教师专业研讨会整合在一起。参与这项服务学习实习和推广计划的本科生将在PI的实验室接受培训，以进行旨在激发对生物技术兴趣的课堂实验，并以简单但科学的方式解释基本概念。该计划旨在让来自不同背景的K-12学生接触大学经历，并通过促进与大学生的接触提供榜样。

With the support of the Organic Dynamics Program in the Chemistry Division, Professor Giovanna Ghirlanda of Arizona State University will design and study novel functional membrane proteins with tunable properties. As a first step in this direction, a heme-binding site has been engineered into a natural membrane protein, Glycophorin A, which forms stable dimers by the association of its transmembrane helical domain. The initial design, ME1, binds ferric protoporphyrin IX with high affinity, exhibits a redox potential within the range of natural membrane heme-binding proteins, and shows nascent catalytic activity. During the funding period this system will be further investigated and serve as the template for the design of a large library of mutants explicitly designed to optimize heme binding and controlling the redox potential. The library will be used to analyze the importance of the heme environment, its hydrophobicity and the presence of aromatic residues in determining the midpoint potential and the binding affinity within the context of the ME1 scaffold. Building on these results ME1 will be evolved into an artificial enzyme for monooxigenase and peroxidase reactivity.The Organic and Macromolecular Chemistry Program supports Professor Giovanna Ghirlanda of Arizona State University whose research on membrane-associated proteins is a relatively untapped resource for applications as biosensors and biocatalysts. A deeper understanding of the relationship between protein structure and function in the membrane environment will permit the preparation of artificial miniaturized proteins tailored to specific biological and non-biological applications. Dr. Ghirlanda will partner with an existing program at ASU to implement a comprehensive approach that integrates classroom demonstrations in K-12 schools, undergraduate training, and specialized workshops for teachers. Undergraduates participating in this service learning internship and outreach program will be trained in the PI's lab to perform classroom experiments designed to spark an interest in biotechnology and to explain the underlying concepts in a simple but scientifically sound manner. The program aims at exposing K-12 students from diverse backgrounds to the college experience and providing role models by facilitating contact with college students.