Abstract
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 P31 and a new unit cell, amenable for room-temperature X-ray diffraction studies. We co-crystallized hAChE in P31 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 P31 space group structures appear similar to those previously obtained in the different P3121 unit cell. Successful solution of the new room temperature 3.2 Å resolution structure of BW284c51*hAChE complex from large P31 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.
Original language | English |
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Article number | 108698 |
Journal | Chemico-Biological Interactions |
Volume | 309 |
DOIs | |
State | Published - Aug 25 2019 |
Funding
X-ray crystallographic data presented in this report are derived from work performed at Argonne National Laboratory, Structural Biology Center (SBC) at the Advanced Photon Source. Use of the Advanced Photon Source, an Office of Science User Facility operated for the U.S. Department of Energy (DOE) Office of Science by Argonne National Laboratory, was supported by the USDOE under Contract No. DE-AC02-06CH11357 . The Office of Biological and Environmental Research supported research at the Center for Structural Molecular Biology (CSMB) at ORNL using facilities supported by the Scientific User Facilities Division , Office of Basic Energy Sciences, U.S. Department of Energy . This research was supported by the CounterACT Program, National Institutes of Health Office of the Director (NIH OD) , and the National Institute of Neurological Disorders and Stroke , [Grant Numbers U01 NS083451 and R21 NS098998 ]. This research was supported by the CounterACT Program, National Institutes of Health Office of the Director (NIH OD), and the National Institute of Neurological Disorders and Stroke, [Grant Numbers U01 NS083451 and R21 NS098998].X-ray crystallographic data presented in this report are derived from work performed at Argonne National Laboratory, Structural Biology Center (SBC) at the Advanced Photon Source. Use of the Advanced Photon Source, an Office of Science User Facility operated for the U.S. Department of Energy (DOE) Office of Science by Argonne National Laboratory, was supported by the USDOE under Contract No. DE-AC02-06CH11357. The Office of Biological and Environmental Research supported research at the Center for Structural Molecular Biology (CSMB) at ORNL using facilities supported by the Scientific User Facilities Division, Office of Basic Energy Sciences, U.S. Department of Energy.
Keywords
- 9-Aminoacridine
- BW284c51
- Donepezil
- Human acetylcholinesterase
- Room-temperature X-ray structure
- Structure-based drug design