From Fundamental Studies of Metalloproteins to Practical Applications

从金属蛋白的基础研究到实际应用

• 批准号: 10580467
• 负责人: OLGA MAKHLYNETS
• 金额: $ 32.81万
• 依托单位: SYRACUSE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-15 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10580467
• 关键词: Active Sites; Address; Affinity; Binding; Biological; Chemical Weapons; Chemicals; Consensus; Cysteine; Defense Mechanisms; Development; Environment; Enzymes; Evolution; Family; Genes; Human; Human Activities; Hydrogen Peroxide; Ions; Iron; Lignin; Manganese; Metalloproteins; Metals; Methane; Methanol; Modeling; Molecular; Nature; Nucleotides; Nutritional; Organism; Oxidants; Oxidation-Reduction; Oxides; Oxygen; Pathogenicity; Pathway interactions; Pesticides; Phenols; Pneumonia; Process; Property; Protein Family; Proteins; Reaction; Reporting; Ribonucleotide Reductase; Scaffolding Protein; Site; Starvation; Streptococcus sanguis; Structure; Structure-Activity Relationship; Study models; System; Testing; Time; Work; biomaterial compatibility; catalyst; chemical reaction; cofactor; depolymerization; design; experimental study; extracellular; fascinate; human pathogen; improved; insight; metalloenzyme; oxidation; practical application; preference; protein function; rational design; remediation; response; sensor; undergraduate student; uptake

Metalloenzymes are capable of efficiently tuning the properties of a metal ion to catalyze very difficult chemical transformations. Yet the determinants of what guides the evolution of protein function still remain not fully understood. This lack of understanding of the subtle detail of interactions that determine enzymatic function limits our ability to rationally design catalysts. We will test why the same family of enzymes uses different metallocofactors. Metalloenzyme NrdF belongs to a class of ribonucleotide reductases (RNR), essential enzymes found in all organisms to catalyze the conversion of nucleotides to deoxynucleotides. RNRs rely on metals to oxidize a conserved cysteine in the active site into a thiyl radical, which then initiates nucleotide reduction. Several different classes of RNRs have been identified in various organisms and, interestingly, despite a remarkable conservation of the overall catalytic pathway, the enzyme can utilize different metals to achieve it. We have recently shown that dimanganese center in class Ib RNRs of pathogenic organisms performs the same task as the diiron center in humans. Moreover, other class I RNR enzymes utilize diiron and mixed iron- manganese centers for function. The molecular and biological determinants of this metal preference still remain unknown and present a major unanswered question in the field. To address this question, we devised three specific aims. Aim 1. Elucidation of the origins of specific metalation in RNR. Using class Ib RNR from Streptococcus sanguinis (Ss) we will address the fundamental question of how correct metalation of enzymes is controlled. Aim 2. Design of functional RNR models. We will use a stable and simple protein model of RNR (DFsc) to bind various metal ions and generate catalysts for practically useful reactions. This aim will give simple, inexpensive and biocompatible protein catalysts for redox transformations and pesticide/chemical weapons remediation in the environment, that could be easily handled in the applied setting. Aim 3. Structural characterization of metalloproteins. Here we will test how metal ions influence the structure of the protein. The work proposed in this aim will validate and correlate the studies on the natural enzymes.

金属酶能够有效地调整金属离子的性能以催化非常困难的化学物质 转型。然而，指导蛋白质功能演变的决定因素仍然不完全 理解。缺乏对确定酶函数限制的相互作用细节的理解 我们合理设计催化剂的能力。我们将测试为什么同一酶使用不同的酶 金属生产子。金属酶NRDF属于一类核糖核苷酸还原酶（RNR），必不可少的 在所有生物体中发现的酶可催化核苷酸向脱氧核苷酸的转化。 RNR依靠 金属将活性位点中的保守半胱氨酸氧化成硫基质，然后启动核苷酸 减少。在各种生物体中已经确定了几种不同类别的RNR，有趣的是 酶是整体催化途径的显着保护，可以利用不同的金属实现它。 我们最近表明，致病生物的IB类RNR中的Dimanganese中心表现相同 任务是人类的二龙中心。此外，其他I类RNR酶也利用二龙和混合铁 - 锰的功能中心。该金属偏好的分子和生物决定因素仍然 保持未知，并在该领域提出一个主要的未解决问题。为了解决这个问题，我们 设计了三个特定目标。目的1。阐明RNR中特定金属的起源。使用IB类 来自Sanguinis链球菌（SS）的RNR我们将解决如何正确金属化的基本问题 酶受到控制。目标2。功能RNR模型的设计。我们将使用稳定而简单的蛋白质模型 RNR（DFSC）的结合，以结合各种金属离子并生成催化剂，以实现实际有用的反应。这个目标 提供简单，廉价和生物相容性的蛋白质催化剂，用于氧化还原转化和农药/化学 在环境中可以轻松处理环境中的武器修复。目标3。结构 金属蛋白的表征。在这里，我们将测试金属离子如何影响蛋白质的结构。 在此目标中提出的工作将验证和关联自然酶的研究。