Overlay independent X-ray data analysis for enhanced oxime OP-ChE reactivation

增强肟OP-ChE再活化的重叠独立X射线数据分析 项目摘要。大量平民和军人易受 有机磷酸盐（OP）暴露仍然是当今的不幸的危险。两者都是神经毒剂 和OP基农药是现实的和令人震惊的重大危害。最近恐怖的巨大神经 叙利亚的毒剂中毒事件和全球每天与OP基农药有关的死亡人数 暴露表明，目前采取的对抗OP的措施普遍缺乏适当性 中毒重点是可负担性、可获得性和有效性。超过200个乙酰胆碱酯酶和 在过去的25年里，沉积在PDB中的BChE结构揭示了异常一致的蛋白质构象， 不仅是蛋白质主链，而且是大多数氨基酸侧链。这种明显的缺乏 在低温和固态结晶状态下记录的构象柔性，表现为 与极高的催化输出相反，特别是在长而窄的AChE活性中心中 峡谷. AChE和BChE蛋白质的催化必需的、热驱动的“呼吸”波动 骨干一直被怀疑和提出，但从未得到明确的证明 实验性的特别是对于肟再活化反应，骨架波动似乎是必不可少的，这是由于 在狭窄的活动中心空间中高度限制对共价连接的OP的访问， 共价结合的OP缀合物。为了设计加速的、有效的再活化剂分子，关键是 知道活性中心哪些部分具有移动和促进亲核肟化的结构倾向 以几何优化的方式接近磷原子。这一信息载于 数百个解决和沉积的胆碱酯酶结构，我们的目标是提取它到一个有用的形式。 我们期望检测到规模较小但性质上系统性的结构性波动。我们最初 人工分析证实了这些预期。

Overlay independent X-ray data analysis for enhanced oxime OP-ChE reactivation Project Summary. Vulnerability of large civilian and military populations to organophosphate (OP) exposure remains an unfortunate peril of the present day. Both, nerve agent OPs and OP based pesticides are realistic and alarmingly significant hazards. Recent horrifying massive nerve agent poisoning episodes in Syria and daily worldwide fatalities associated with OP based pesticide exposures reveal general lack of appropriateness of currently employed measures for counteracting OP intoxications. The emphasis is on affordability, availability and efficacy. More than two hundred AChE and BChE structures deposited in PDB in the past 25 years reveal unusually consistent protein conformations, not only of protein backbones but also of majority of amino acid side chains. This apparent absence of conformational flexibility registered at the low temperature and in the solid crystaline state, appears as a contrast to an extreemely high catalytic output, in particular in the long and narrow AChE active center gorge. Catalytically essential, thermally driven, „breathing“ fluctuations of the AChE and BChE protein backbones have long been suspected and proposed but have never been clearly demonstrated experimentally. In particular for oxime reactivation reactions backbone fluctuations appear essential due to highly restricted access to covalently attached OPs in the narrow actice center space impacted further by covalently bound OP conjugates. For design of an accelerated, efficient reactivator molecule it is critical to know what parts of active center have structural predisposition to move and facilitate nucleophilic oxime approach to the phosphorus atom in geometrically optimal fashion. This information is encripted in hundreds of resolved and deposited ChE structures and our goal is to extract it to an useful form. We expect to detect structural fluctuations small in magnitude yet systematic in nature. Our initial manual analyses confirm those expectations.