Biodynamics: Vibrational Echo Correlation Spectroscopy

生物动力学：振动回波相关光谱

• 批准号: 6771506
• 负责人: MICHAEL D FAYER
• 金额: $ 28.2万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6771506
• 关键词: active sites; allosteric site; binding sites; cytochrome c; hemocyanin; hemoglobin; hemoprotein; infrared spectrometry; intermolecular interaction; lysozyme; metalloproteins; method development; molecular dynamics; myoglobin; nanotechnology; protein denaturation; protein protein interaction; protein structure; structural biology; time resolved data; vibration; water; water environment

Research is proposed to study dynamics and structure of biologically important systems including heme proteins, proteins with multiple metal centers, water in nanoscopic environments, and water-protein interactions in nanoscopic environments. Building on our current work and recent experimental and theoretical advances make possible the application of Vibrational Echo Correlation Spectroscopy, a powerful new approach to the study of these problems. Vibrational echo correlation spectroscopy with full phase information will be used to directly examine the structural degrees of freedom and dynamical interactions of biologically important systems in a manner that is akin to multidimensional NMR. The new research expands our currently successful application of multidimensional vibrational echo methods. Extending work on myoglobin-CO, mutants will be used to unravel structural dynamics throughout the protein. Experiments on hemoglobin-CO will relate the allosteric affect to protein dynamics at the active site and examine the temperature dependence of structural evolution. Cytochrome c mutant M80A, where the axial methionine residue is replaced with an alanine binds both CO and CN-, in the Fe+2 and Fe+3 oxidation states, respectively. This mutant will be studied to address fundamental dynamical differences that occur in the protein when the oxidation state of the heme group is changed. Proteins with multiple Cu centers will be studied. First, hemocyanin will be studied with CO bound at the active site, and insights into the active site protein dynamics gained from studies of hemocyanin will be applied to other binuclear copper proteins such as tyrosinase. The hemocyanin experiments are a precursor to the study of trinuclear Cu proteins such as laccase, ascorbic oxidase, and ceruloplasmin. Multiple azide anions will be bound to these electronically coupled Cu centers to study the dynamics and vibrational mode coupling between the azide ligands. Vibrational echo correlation spectroscopy provides a new type of analytical tool for the study of coupled metal centers that will also be applied to the several multiple metal centers in carbon monoxide dehydrogenasetacetyI-CoA synthase (CODH/ACS). Building on recent fundamentally new studies of water dynamics, the biologically important issue of the dynamics of water in nanoscopic environments will be studied. In addition, water-protein dynamical interactions and water-protein hydrogen bond dynamics in nanoscopic water environments will be examined.

拟研究生物学重要系统的动力学和结构，包括血红素蛋白、具有多个金属中心的蛋白质、纳米环境中的水以及纳米环境中的水-蛋白质相互作用。基于我们当前的工作以及最近的实验和理论进展，振动回波相关光谱的应用成为可能，这是研究这些问题的一种强大的新方法。具有全相位信息的振动回波相关光谱将用于以类似于多维核磁共振的方式直接检查生物学重要系统的结构自由度和动态相互作用。新的研究扩展了我们目前的 多维振动回波方法的成功应用。突变体将用于扩展肌红蛋白-CO 的研究，以揭示整个蛋白质的结构动力学。血红蛋白-CO 实验将把变构影响与活性位点的蛋白质动力学联系起来，并检查结构演化的温度依赖性。细胞色素 c 突变体 M80A，其中轴向甲硫氨酸残基被丙氨酸取代，分别在 Fe+2 和 Fe+3 氧化态下结合 CO 和 CN-。我们将研究这种突变体，以解决当血红素基团的氧化态改变时蛋白质中出现的基本动力学差异。将研究具有多个铜中心的蛋白质。首先，将研究血蓝蛋白与活性位点结合的CO，并且从血蓝蛋白研究中获得的对活性位点蛋白质动力学的见解将应用于其他双核铜蛋白，例如酪氨酸酶。血蓝蛋白实验是漆酶、抗坏血酸氧化酶和铜蓝蛋白等三核铜蛋白研究的先驱。多个叠氮阴离子将与这些电子耦合的铜中心结合，以研究叠氮配体之间的动力学和振动模式耦合。振动回波相关光谱为耦合金属中心的研究提供了一种新型分析工具，也将应用于多种 一氧化碳脱氢乙酰辅酶 A 合酶 (CODH/ACS) 中的多个金属中心。在最近对水动力学的全新研究的基础上，将研究纳米环境中水动力学的生物学重要问题。此外，还将研究纳米水环境中的水-蛋白质动力学相互作用和水-蛋白质氢键动力学。