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

基本信息

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
项目类别：
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

项目摘要

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）：我们在此提出一项尖端生物物理研究，旨在确定神经毒剂肟再激活中结构上施加的限制因素和农药有机磷（OP）抑制人乙酰胆碱酯酶（hAChE）。我们将重点关注溶液中 hAChE 蛋白结构域运动的特征，这对于促进重新激活至关重要，以及参与共价结合磷的亲核取代的 hAChE 活性中心的精确质子结构。该研究将使用尖端技术，根据平衡状态下的 X 射线和中子散射来表征溶液中 hAChE 的再激活相关分子运动，并在 ps 到 min 间隔的时间范围内进行解析。高分辨率联合室温下 hAChE 晶体上的 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单独的天然、OP 缀合和老化的 hAChE 以及与选定的结构多样的肟重激活剂的复合物的比较分析，旨在确定肟的结构特征，以克服结构动力学和结构所施加的限制。 HAChE 活性中心的质子分布。获得的信息与先进的计算分析和模拟相结合将用于设计 OP-hAChE 的原型加速肟重激活剂。原型加速再激活剂的结构将可使用我们与拉霍亚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进行合成和精制。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Rational design, synthesis, and evaluation of uncharged, "smart" bis-oxime antidotes of organophosphate-inhibited human acetylcholinesterase.

有机磷抑制人乙酰胆碱酯酶的不带电荷的“智能”双肟解毒剂的合理设计、合成和评估。

DOI：
发表时间：
2020
期刊：
影响因子：
0
作者：
Gorecki, Lukas;Gerlits, Oksana;Kong, Xiaotian;Cheng, Xiaolin;Blumenthal, Donald K;Taylor, Palmer;Ballatore, Carlo;Kovalevsky, Andrey;Radić, Zoran
通讯作者：
Radić, Zoran

Productive reorientation of a bound oxime reactivator revealed in room temperature X-ray structures of native and VX-inhibited human acetylcholinesterase.

天然和 VX 抑制的人乙酰胆碱酯酶的室温 X 射线结构揭示了结合肟重激活剂的有效重新定向。

DOI：
发表时间：
2019
期刊：
影响因子：
0
作者：
Gerlits, Oksana;Kong, Xiaotian;Cheng, Xiaolin;Wymore, Troy;Blumenthal, Donald K;Taylor, Palmer;Radić, Zoran;Kovalevsky, Andrey
通讯作者：
Kovalevsky, Andrey

A new crystal form of human acetylcholinesterase for exploratory room-temperature crystallography studies.

用于探索性室温晶体学研究的人乙酰胆碱酯酶的新晶体形式。

DOI：
发表时间：
2019-08-25
期刊：
影响因子：
5.1
作者：
Gerlits, Oksana;Ho, Kwok;Cheng, Xiaolin;Blumenthal, Donald;Taylor, Palmer;Kovalevsky, Andrey;Radić, Zoran
通讯作者：
Radić, Zoran

Structural and dynamic effects of paraoxon binding to human acetylcholinesterase by X-ray crystallography and inelastic neutron scattering.

通过 X 射线晶体学和非弹性中子散射研究对氧磷与人乙酰胆碱酯酶结合的结构和动力学效应。

DOI：
发表时间：
2022-11-03
期刊：
影响因子：
0
作者：
Gerlits, Oksana;Fajer, Mikolai;Cheng, Xiaolin;Blumenthal, Donald K.;Radic, Zoran;Kovalevsky, Andrey
通讯作者：
Kovalevsky, Andrey