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：
10238801
负责人：
Matthew Chalkley
金额：
$ 6.6万
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 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10238801
关键词：
Active Sites Affinity Amino Acids Anabolism Antibiotics Binding Binding Proteins Bioavailable 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：金属酶以保持速率和选择性进行化学转化嫉妒合成化学家。根据定义，这些转变利用了地球丰富的过渡金属和环保试剂。此外，有些金属酶使用专业的辅助因子，许多人能够使用相对有限的天然配体由氨基酸提供。确实，在许多情况下用于促进各种机械不同的转换，为证据提供证据蛋白质环境的二级和三级结构的重要机制。理解结构功能原理的一种方法是从头设计金属酶从划痕。本文中，我们利用从头蛋白质设计，使我们能够系统地改变生物学上重要的，弹性反应周围的本地环境中级，铁超氧。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物理上重要的C – H激活反应在单核，非血红素Fe中心。 C – H激活反应是从结构功能的角度特别兴趣，因为他们的成功已显示高度依赖底物定位，从而提供了对我们的能力的敏感测试从头设计装订口袋。提高我们设计受控装订口袋的能力将为从头蛋白的许多应用打开可能性。拟议的研究将主要可以使用光谱法来表征电子结构和不同物种的热化学。这些研究将通过结构性完成来自多核NMR光谱和X射线晶体学的信息。结果将代表蛋白质设计的突破，对基本理解的影响金属蛋白如何利用简单的配体套件生成和线束反应性中间体为了实现化学挑战转型。