Design and Evolution of Metal-Based Functions in Supramolecular Protein Scaffolds

超分子蛋白支架中金属基功能的设计与演化

• 批准号: 10221740
• 负责人: Faik Akif Tezcan
• 金额: $ 31.29万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 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 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10221740
• 关键词: Address; Affinity; Amides; Architecture; Automobile Driving; Binding; Biochemistry; Bioinorganic Chemistry; Biology; Biomimetics; Catalysis; Cell Respiration; Cell physiology; Chemistry; Complex; Cytochromes; Dioxygen; Directed Molecular Evolution; Elements; Engineering; Environment; Esters; Evolution; Genetic Transcription; Geometry; Goals; Health; Human; Hydrogen Bonding; Hydrolase; Hydrolysis; Ions; Knowledge; Lead; Mediating; Metal Ion Binding; Metalloproteins; Metals; Methods; Mutation; Natural Products; Organism; Oxidation-Reduction; Pathway interactions; Peptides; Phosphoric Monoester Hydrolases; Property; Protein Engineering; Proteins; Reaction; Research; Research Project Grants; Respiration; Scaffolding Protein; Series; Spectrum Analysis; Structure; Structure-Activity Relationship; Surface; Testing; Thermodynamics; Time; Transition Elements; amidase; base; beta-Lactamase; design; design and construction; dimer; esterase; flexibility; improved; knowledge of results; metal complex; metalloenzyme; microbial; neurotransmission; novel; novel strategies; oxidation; polypeptide; programs; protein structure; rational function; scaffold; self assembly

Metalloproteins carry out many cellular functions that are central to biology and human health. While our knowledge of how metalloproteins function has grown immensely thanks to technological advances, we still possess only a superficial understanding of the interplay between protein structure/dynamics and metal coordination/reactivity. As a result, it has not yet been possible a) to predict the functional mechanism of metalloproteins simply by looking at their structures, b) to emulate or improve upon the structures and functions of metalloproteins by de novo design, and c) to understand how complex bioinorganic functions may have emerged on simple peptide/protein scaffolds during natural evolution. The overarching goal of the proposed research program is to address these three challenges by designing and constructing protein scaffolds with increasingly more complex metal-based functions from scratch. Inspired by a hypothetical pathway for the natural evolution of metalloproteins, we have recently developed new approaches to metalloprotein design in which monomeric proteins are templated by metal ions to form novel supramolecular assemblies. The interfaces of these evolutionarily naïve complexes are then engineered and evolved to create self-standing protein architectures with complex metal-based functions. In the proposed research, we will further develop these “metal-templated protein design” strategies by capitalizing on two new protein scaffolds developed in our lab (DiCyt and TriCyt), which provide easy access to diverse metal coordination geometries, secondary-sphere environments and global properties (tunable structures, oligomeric states, flexibility/rigidity) that are difficult to attain with other protein design strategies. We will use DiCyt and TriCyt scaffolds to build metalloprotein assemblies for stable and selective coordination of first-row transition metal ions (Specific Aim 1), for challenging ester, amide and phospho-ester bond hydrolysis reactions (Specific Aim 2), and for redox catalytic reactions involving dioxygen binding and activation (Specific Aim 3). These efforts will not only uncover fundamental structure-function relationships that govern diverse metalloprotein activities, but also lead to better understanding of how bioinorganic complexity emerges in simple protein scaffolds.

金属蛋白执行许多对生物学和人类健康至关重要的细胞功能。虽然我们的 由于技术进步，关于金属蛋白如何发挥作用的知识已大大增长，但我们仍然 对蛋白质结构/动力学与金属之间的相互作用仅具有肤浅的了解 协调性/反应性。因此，目前还不可能 a) 预测其功能机制 只需观察金属蛋白的结构，b) 即可模拟或改进其结构和功能 通过从头设计来研究金属蛋白，以及 c) 了解复杂的生物无机功能如何发挥作用 在自然进化过程中出现在简单的肽/蛋白质支架上。拟议的总体目标 研究计划是通过设计和构建蛋白质支架来解决这三个挑战 从头开始实现越来越复杂的金属功能。受到自然的假设途径的启发 金属蛋白的进化，我们最近开发了金属蛋白设计的新方法，其中 单体蛋白质以金属离子为模板形成新型超分子组装体。的接口 然后，这些进化上幼稚的复合物被设计和进化，以产生独立的蛋白质 具有复杂的基于金属的功能的架构。在拟议的研究中，我们将进一步开发这些 利用我们实验室开发的两种新蛋白质支架的“金属模板蛋白质设计”策略 （DiCyt 和 TriCyt），可轻松获得不同的金属配位几何形状、次级球体 环境和全局特性（可调结构、寡聚状态、灵活性/刚性）是很难 与其他蛋白质设计策略一起实现。我们将使用 DiCyt 和 TriCyt 支架来构建金属蛋白 用于第一行过渡金属离子的稳定和选择性配位的组件（具体目标 1），以应对具有挑战性的问题 酯、酰胺和磷酸酯键水解反应（具体目标 2），以及氧化还原催化反应 涉及双氧结合和激活（具体目标 3）。这些努力不仅会揭示根本性的 控制不同金属蛋白活性的结构-功能关系，也有助于更好地理解 生物无机复杂性如何在简单的蛋白质支架中出现。