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: 9632884
• 负责人: Zoran Radic
• 金额: $ 3.16万
• 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/9632884
• 关键词: Acetylcholine; Acetylcholinesterase; Ache; Active Sites; Address; Antidotes; Catalysis; Cholinesterases; Collaborations; Complex; Computer Analysis; Computer Simulation; Conflict (Psychology); Coupled; Crystallization; Data; Dependence; Development; Dimensions; Drug Design; Enzymes; Equilibrium; Equipment and supply inventories; Event; Future; Generations; Human; Hydration status; Hydrogen; Joints; Knowledge; Laboratories; Ligands; Location; Mediating; Methods; Molecular; Molecular Conformation; Monitor; Motion; Mus; Neurotransmitters; Neutron Diffraction; Neutrons; Organophosphates; Oximes; Oxygen; Pesticides; Phosphorus; Positioning Attribute; Process; Property; Proteins; Protons; Reaction; Refractory; Research Institute; Research Personnel; Resolution; Roentgen Rays; Scheme; Side; 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; public health relevance; screening; three dimensional structure; Acetylcholine; Acetylcholinesterase; Ache; Active Sites; Address; Antidotes; Catalysis; Cholinesterases; Collaborations; Complex; Computer Analysis; Computer Simulation; Conflict (Psychology); Coupled; Crystallization; Data; Dependence; Development; Dimensions; Drug Design; Enzymes; Equilibrium; Equipment and supply inventories; Event; Future; Generations; Human; Hydration status; Hydrogen; Joints; Knowledge; Laboratories; Ligands; Location; Mediating; Methods; Molecular; Molecular Conformation; Monitor; Motion; Mus; Neurotransmitters; Neutron Diffraction; Neutrons; Organophosphates; Oximes; Oxygen; Pesticides; Phosphorus; Positioning Attribute; Process; Property; Proteins; Protons; Reaction; Refractory; Research Institute; Research Personnel; Resolution; Roentgen Rays; Scheme; Side; 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; public health relevance; 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）：我们在这里提出了一项尖端生物物理研究，旨在鉴定神经剂和 农药有机磷酸盐（OP）抑制人乙酰胆碱酯酶（HACHE）。我们将专注于表征Hache蛋白结构域运动在促进重新激活至关重要的解决方案中，以及与共价共轭磷的亲核取代的Hache活性中心的精确质子结构。该研究将使用尖端技术来表征基于X射线和中子散射在平衡中的溶液中Hache的重新激活相关的分子运动，并在从PS到最小间隔的时间内解决。高分辨率关节 室温下Hache晶体上的X射线/中子衍射将用于解决Hache活动中心中质子的精确位置和完整的水分子。散射和衍射研究都将依赖于对天然，运行偶联和单独老化的Hache的比较分析，并且与选定的结构上多样的棒棒棒激活剂具有复杂性，并将旨在识别Oximes的结构特征，以实现Hache Active Center中的结构动力学和Proton分布所施加的过度限制。获得的信息以及高级计算分析和仿真将用于设计OP-HACHE的原型加速Oxime Reactivator。使用现有的药物设计管道与La Jolla的Scripps研究所的研究人员合作开发的原型加速反抗器的结构将用于合成和完善。