O2-Reactivity in De Novo Designed Mononuclear, Non-Heme Fe Proteins

De Novo 设计的单核非血红素铁蛋白中的 O2 反应性

基本信息

批准号：
10468710
负责人：
Matthew Chalkley
金额：
$ 6.26万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2023-07-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10468710
关键词：
Active Sites Affinity Amino Acids Anabolism Antibiotics Binding Binding Proteins Biochemical Biological Biological Models Biology Characteristics Charge Chemicals Complement Complex Computing Methodologies DNA Dioxygen Electrons Environment Feedback Future Health Heme Histidine Human Hydrogen Bonding Hydrogen Peroxide Hydroxylation Iron Keto Acids Length Ligands Location Maintenance Mediating Metalloproteins Metals Mononuclear Multinuclear NMR NMR Spectroscopy Nature Optics Oxidation-Reduction Phase Physiological Planet Earth Porphyrins Positioning Attribute Post-Translational Protein Processing Property Protein Engineering Proteins RNA Reaction Reagent Specificity Spectrum Analysis Structure Structure-Activity Relationship Techniques Tertiary Protein Structure Testing Time Transition Elements Work X-Ray Crystallography carboxylate chemical bond cofactor design electronic structure improved insight interest metalloenzyme novel strategies oxidation repaired small molecule success

项目摘要

Project Summary/Abstract: Metalloenzymes perform chemical transformations with rates and selectivities that remain the envy of synthetic chemists. By definition these transformations utilize earth-abundant transition metals and environmentally friendly reagents. Furthermore, while some metalloenzymes utilize specialized cofactors, many are able to achieve these transformations using the relatively limited natural ligand set provided by the amino acids. Indeed, in many cases a single coordination motif is used to promote a variety of mechanistically distinct transformations providing evidence for the important of the secondary and tertiary structure of the protein environment for dictating reaction mechanism. One approach to understanding the structure-function principles is to de novo design metalloenzymes from scratch. Herein we exploit de novo protein design to allow us to systematically alter the local environment around a biologically important, ambiphilic reaction intermediate, the ferric superoxo. We then seek to utilize this understanding and the newfound ability to design specific small molecule binding proteins to explore physiologically important C– H activation reactions at a mononuclear, non-heme Fe center. C–H activation reactions are of particular interest from a structure-function perspective because their success has been shown to be highly dependent on substrate positioning, thereby providing a sensitive test of our ability to de novo design binding pockets. Improving our ability to design controlled binding pockets would open the possibility for many applications of de novo proteins. The proposed studies will primarily be achieved using optical spectroscopy to characterize the electronic structure and thermochemistry of the different species. These studies will be complemented by structural information derived from multinuclear NMR spectroscopy and X-ray crystallography. The results would represent a breakthrough in protein design with implications for fundamental understanding of how metalloproteins utilize simple ligand sets to generate and harness reactive intermediates for achieving chemically challenging transformations.

项目摘要/摘要：金属酶进行化学转化的速率和选择性仍然保持在根据定义，这些转化利用了地球上丰富的转变。此外，虽然一些金属酶利用金属和环保试剂。专门的辅助因子，许多人能够使用相对有限的方法来实现这些转化事实上，在许多情况下，由氨基酸提供的天然配体组。用于促进各种机械上不同的转变，为以下事实提供证据：蛋白质环境的二级和三级结构对于决定反应的重要性理解结构功能原理的一种方法是从头设计。在此，我们利用从头开始的蛋白质设计，使我们能够系统地改变具有重要生物学意义的双亲反应周围的局部环境然后我们寻求利用这种理解和新发现。能够设计特定的小分子结合蛋白来探索生理上重要的 C– 单核、非血红素 Fe 中心的 H 活化反应为 C-H 活化反应。从结构功能的角度特别感兴趣，因为它们的成功已被证明高度依赖于基材的定位，从而为我们的能力提供了一个敏感的测试从头设计装订口袋将提高我们设计受控装订口袋的能力。拟议的研究将主要为从头蛋白质的许多应用提供可能性。可以使用光谱来表征电子结构和不同物种的热化学研究将得到结构的补充。来自多核核磁共振波谱和 X 射线晶体学的信息。将代表蛋白质设计的突破，并对基本理解产生影响金属蛋白如何利用简单的配体组来生成和利用反应中间体实现具有化学挑战性的转化。