Accelerated AChE Reactivator Design by Mechanistic Neutron Scattering Studies

通过机械中子散射研究加速乙酰胆碱酯酶再激活器设计

• 批准号: 8892277
• 负责人: Zoran Radic
• 金额: $ 67.02万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-15 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8892277
• 关键词: Acetylcholine; Acetylcholinesterase; Active Sites; Address; Antidotes; Binding; Catalysis; Cholinesterases; Collaborations; Complex; Computer Analysis; Computer Simulation; Conflict (Psychology); Coupled; Data; Dependence; Development; Dimensions; Drug Design; Electronics; Enzymes; Equilibrium; Equipment and supply inventories; Event; Future; Generations; Health; Human; Hydration status; Hydrogen; Joints; Knowledge; Laboratories; Ligands; Location; Mediating; Methods; Molecular; Monitor; Motion; Mus; Neurotransmitters; Neutron Diffraction; Neutrons; Organophosphates; Oximes; Oxygen; Pesticides; Phosphorus; Positioning Attribute; Process; Property; Proteins; Protons; Reaction; Refractory; Relative (related person); Research Institute; Research Personnel; Resolution; Roentgen Rays; Scheme; Side; Solutions; Solvents; Spectrometry; Structure; System; Techniques; Technology; Temperature; Tertiary Protein Structure; Testing; Time; Triad Acrylic Resin; Universities; Utah; Water; X-Ray Crystallography; aged; base; biophysical analysis; comparative; design; improved; insight; instrument; ionization; millisecond; nerve agent; next generation; novel strategies; nucleophilic substitution; protonation; screening; three dimensional structure

DESCRIPTION (provided by applicant): We propose here a cutting edge biophysical study aimed at identifying structurally imposed limiting factors in oxime reactivation of nerve agent and pesticide organophosphate (OP) inhibited human acetylcholinesterase (hAChE). We will focus on characterizing hAChE protein domain motions in solution critical for facilitating reactivation a well as on precise proton structure in the hAChE active center involved in the nucleophilic substitution of covalently conjugated phosphorus. The study will use cutting edge techniques to characterize reactivation related molecular motions of hAChE in solution based on X-ray and neutron scatter in equilibrium and resolved in time from ps to min intervals. High resolution joint X-ray/neutron diffraction on hAChE crystals at room temperature will be used to resolve precise positions of protons and complete water molecules in the hAChE active center. Both scattering and diffraction studies will rely on comparative analysis of native, OP-conjugated, and aged hAChE alone, and in complex with selected structurally diverse oxime reactivators and will be aimed at identifying structural features of oximes optimal for overcoming limitations imposed by structural dynamics and proton distribution in the hAChE active center. The acquired information coupled with advanced computational analysis and simulation will be used to design prototypic accelerated oxime reactivator of OP-hAChE. The structures of prototypic accelerated reactivators will be made available for synthesis and refinement using our existing drug design pipeline developed in collaboration with researchers of The Scripps Research Institute in La Jolla.

描述（由申请人提供）：我们在此提出了一项尖端生物物理学研究，旨在确定神经毒剂肟再活化中结构上强加的限制因素， 有机磷农药（OP）抑制人乙酰胆碱酯酶（hAChE）。我们将专注于表征hAChE蛋白质结构域的运动在溶液中的关键促进再活化以及精确的质子结构中的hAChE活性中心参与的亲核取代共价共轭磷。这项研究将使用尖端技术来表征再活化相关的分子运动的hAChE在溶液中的基础上的X射线和中子散射的平衡和解决的时间从ps到分钟的间隔。高分辨率接头 在室温下对hAChE晶体的X射线/中子衍射将用于解析hAChE活性中心中质子和完整水分子的精确位置。散射和衍射研究都将依赖于对单独的天然、OP缀合和老化的hAChE以及与选定的结构不同的肟再活化剂的复合物的比较分析，目的是确定最适合克服结构动力学和质子所施加的限制的肟的结构特征。hAChE活性中心的质子分布。所获得的信息加上先进的计算分析和模拟将用于设计原型加速肟活化剂的OP-hAChE。原型加速再活化剂的结构将可用于合成和改进，使用我们与位于拉霍亚的斯克里普斯研究所的研究人员合作开发的现有药物设计管道。