Minority Supplement for GM111932

GM111932 少数族裔补充品

• 批准号: 9282901
• 负责人: Ronald Koder
• 金额: $ 9.14万
• 依托单位: CITY COLLEGE OF NEW YORK
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-15 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9282901
• 关键词: Active Sites; Affect; Affinity; Amyotrophic Lateral Sclerosis; Binding; Biochemical; Blood Substitutes; Carrier Proteins; Chemicals; Chimera organism; Complex; Cytochrome P450; Dioxygenases; Disease; Distal; Drug Metabolic Detoxication; Electron Transport; Electrons; Electrostatics; Enzymes; Equilibrium; Face; Flavins; Flavoproteins; Future; Health; Heart Diseases; Heme; Heme Iron; Hemoglobin; Histidine; Human; Human Biology; Ions; Ischemic Brain Injury; Learning; Left; Ligand Binding; Ligands; Ligation; Malignant Neoplasms; Medicine; Metabolic; Methods; Minority; Modification; Molecular; Nature; Nitrates; Nitric Oxide; Oxides; Oxidoreductase; Oxygen; Pathway interactions; Penetration; Play; Poison; Property; Protein Binding Domain; Protein Dynamics; Protein Engineering; Proteins; Reaction; Role; Rotation; Scientist; Side; Site; Stroke; Structure; Surface; Technology; Tertiary Protein Structure; Testing; Therapeutic; Thermodynamics; Water; Work; alpha helix; base; cofactor; design; driving force; effective therapy; enzyme therapy; innovation; molecular dynamics; next generation; oxygen transport; phthalate 4,5-dioxygenase; protein structure; semiquinone; signal processing; synthetic enzyme; therapeutic enzyme

DESCRIPTION (provided by applicant): Significance. We aim to determine the essential structural and thermodynamic features which govern enzymatic nitric oxide detoxification. We use a cycle of computational design and biochemical analysis of an artificial nitric oxide dioxygenase (NOD) formed by combining an artificial heme-based oxygen binding protein domain with a flavoprotein reductase domain derived from nature. Use of such a robust, simple protein makes it significantly easier to make both small- and large-scale changes to the protein and positively identify critical features necessary for enzyme function. Nitric oxide plays a central role in many signaling process in human biology, yet due to its degree of chemical reactivity it has also been implicated in a surprising number of serious disorders such as Lou Gehrig's Disease and ischemic brain injury. A superior nitric oxide dioxygenase thus promises to be useful in future treatments of many pathological conditions. Conversely, unwanted NOD activity has produced severe complications in hemoglobin-based blood substitutes, and it is important to learn how to reduce or eliminate NOD activity in these therapeutics without adversely affecting oxygen binding. Innovation. This catalytic construct represents the next generation in protein design, moving design technology from the current focus on simple protein domains with single cofactors to significantly more challenging and sophisticated multidomain structures that more closely resemble the complex assemblies seen in nature. This project has the capacity to dramatically advance two important technologies: hemoglobin-based blood substitutes and enzyme therapeutics. First, lessons learned in this project promise to revitalize the field of hemoglobin-based blood substitutes, enabling both the reengineering of native hemoglobins and the creation of entirely new oxygen transport proteins minimally reactive with nitric oxide while still carrying oxygen. Second, a synthetic enzyme has the potential to transform the field of enzyme therapy because of the many advantages designed enzymes have over their natural counterparts, most importantly the ability to utilize non-natural cofactors better optimized for the target activity and their greatly increased stability over natural protein (53). This project thus represents a new direction in enzyme therapy, and our design pathway is an enabling technology which will be used by us and others in the creation of future enzyme therapeutics. Specific Aims. This work will allow us to answer some important questions about this enzyme: Aim 1. What role does the heme reduction potential play in the nitric oxide dioxygenase reaction? Aim 2. How important are electron transfer dynamics and thermodynamics in this reaction? Aim 3. How do protein dynamics and structure govern NOD function?

描述（由申请人提供）：意义。我们的目的是确定基本的结构和热力学特征，支配酶促一氧化氮解毒。我们使用一个循环的计算设计和生物化学分析的人工一氧化氮双加氧酶（NOD）形成的人工血红素为基础的氧结合蛋白结构域与黄素蛋白还原酶结构域来自自然界。使用这样一个强大的，简单的蛋白质，使它更容易作出小规模和大规模的变化，蛋白质和积极确定酶功能所必需的关键特征。一氧化氮在人类生物学中的许多信号传导过程中起着核心作用，但由于其化学反应性的程度，它也与惊人数量的严重疾病有关，例如Lou Gehrig病和缺血性脑损伤。因此，一种上级一氧化氮双加氧酶有望在许多病理状况的未来治疗中是有用的。相反，不需要的NOD活性已经在基于血红蛋白的血液替代品中产生了严重的并发症，并且重要的是了解如何在这些治疗中减少或消除NOD活性而不对氧结合产生不利影响。创新这种催化构建体代表了蛋白质设计的下一代，将设计技术从目前关注具有单一辅因子的简单蛋白质结构域转移到更具有挑战性和复杂的多结构域结构，这些结构域更接近自然界中看到的复杂组装体。该项目有能力大幅推进两项重要技术：基于血红蛋白的血液替代品和酶疗法。首先，在这个项目中吸取的经验教训有望振兴血红蛋白为基础的血液替代品领域，使天然血红蛋白的再造和创造全新的氧转运蛋白与一氧化氮反应最低，同时仍然携带氧气。第二，合成酶具有改变酶疗法领域的潜力，因为设计的酶与天然酶相比具有许多优点，最重要的是利用非天然辅因子的能力 更好地优化了目标活性，并且其稳定性大大提高，超过天然蛋白质（53）。因此，该项目代表了酶疗法的新方向，我们的设计途径是一种使能技术，将被我们和其他人用于创造未来的酶疗法。具体目标。这项工作将使我们能够回答有关这种酶的一些重要问题：目的1。血红素还原电位在一氧化氮双加氧酶反应中扮演什么角色？目标2.在这个反应中，电子转移动力学和热力学有多重要？目标3。蛋白质动力学和结构如何控制NOD功能？