A new crystal form of human acetylcholinesterase for exploratory room-temperature crystallography studies
A new crystal form of human acetylcholinesterase for exploratory room-temperature crystallography studies
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DOI：
10.1016/j.cbi.2019.06.011
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发表时间：
2019-08-25
影响因子：
5.1
通讯作者：
Radic, Zoran
中科院分区：
文献类型：
--
作者：
Gerlits, Oksana;Ho, Kwok-Yiu;Radic, Zoran
新的药理活性分子的结构导向设计至少部分依赖于基于模板的药物设计的大分子结构的功能相关的准确性。目前，在蛋白质数据库(PDB)中，大约95%的大分子X射线结构是从低温下的衍射实验中获得的。然而，众所周知，在较高的生理温度下，大分子和药物配体的功能相关构象可能不同。本文描述了P3(1)空间群的新型人乙酰胆碱酯酶(AChE)晶体和一种适合于室温X射线衍射研究的新晶胞的发展和性质。我们在P3(1)单位细胞中将HAChE与结合在活性中心峡谷底部的可逆抑制剂9-氨基吖啶以及跨越峡谷全长的抑制剂多奈哌齐(Aricept，E2020)和AChE特异性抑制剂BW284c51共结晶。它们的新低温P3(1)空间群结构看起来与以前在不同的P3(1)21单胞中获得的结构相似。从大的P3(1)晶体中成功地解决了BW284C51*HAChE络合物的新的室温3.2埃分辨结构，使我们能够继续研究低亲和力络合物的室温结构，例如与HAChE结合的肟类复活剂，其中肟类和HAChE中可能存在与温度相关的构象多样性，这可能导致在接近生理温度的条件下更好地基于结构的设计。
Structure-guided design of novel pharmacologically active molecules relies at least in part on functionally relevant accuracy of macromolecular structures for template based drug design. Currently, about 95% of all macromolecular X-ray structures available in the PDB (Protein Data Bank) were obtained from diffraction experiments at low, cryogenic temperatures. However, it is known that functionally relevant conformations of both macromolecules and pharmacological ligands can differ at higher, physiological temperatures. We describe in this article development and properties of new human acetylcholinesterase (AChE) crystals of space group P3(1) and a new unit cell, amenable for room-temperature X-ray diffraction studies. We co-crystallized hAChE in P3(1) unit cell with the reversible inhibitor 9-aminoacridine that binds at the base of the active center gorge in addition to inhibitors that span the full length of the gorge, donepezil (Aricept, E2020) and AChE specific inhibitor BW284c51. Their new low temperature P3(1) space group structures appear similar to those previously obtained in the different P3(1)21 unit cell. Successful solution of the new room temperature 3.2 angstrom resolution structure of BW284c51*hAChE complex from large P3(1) crystals enables us to proceed with studying room temperature structures of lower affinity complexes, such as oxime reactivators bound to hAChE, where temperature-related conformational diversity could be expected in both oxime and hAChE, which could lead to better informed structure-based design under conditions approaching physiological temperature.