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

• 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
• 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

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.

项目概要/摘要： 金属酶进行化学转化的速率和选择性保持不变， 合成化学家的羡慕。根据定义，这些转换利用地球丰富的过渡 金属和环境友好的试剂。此外，虽然一些金属酶利用 由于有专门的辅因子，许多人能够使用相对有限的 由氨基酸提供的天然配体组。事实上，在许多情况下， 是用来促进各种不同的机制转换提供证据， 蛋白质环境的二级和三级结构对于决定反应的重要性 机制理解结构-功能原理的一种方法是重新设计 从零开始在这里，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两亲性反应周围的局部环境， 中间体，超氧铁。然后，我们寻求利用这种理解和新发现的 设计特异性小分子结合蛋白以探索生理学上重要的C- H活化反应在单核，非血红素铁中心。C-H活化反应是 从结构-功能的角度来看，它们特别令人感兴趣，因为它们的成功已经被证明是 高度依赖于基板定位，从而提供了一个敏感的测试我们的能力， 从头设计装订口袋。提高我们设计可控约束口袋的能力， 为从头蛋白质的许多应用开辟了可能性。拟议的研究将主要 使用光谱学来表征电子结构， 不同物种的热化学。这些研究将得到结构性的补充。 来自多核NMR光谱学和X射线晶体学的信息。结果 将代表蛋白质设计的突破， 金属蛋白如何利用简单的配体来产生和利用活性中间体 来实现具有化学挑战性的转化。