期刊论文详细信息
IUCrJ | |
Using cryo-electron microscopy maps for X-ray structure determination | |
Ding, W.1  Zeng, L.2  | |
[1]Institute of Physics, Chinese Academy of Sciences, Beijing 100190, People's Republic of China | |
[2]School of Biomedical Sciences, University of Hong Kong, 21 Sassoon Road, Hong Kong | |
关键词: FSEARCH; IPCAS; ITERATIVE PHASING; CRYO-EM; MODEL BUILDING; STRUCTURE DETERMINATION; CRYO-ELECTRON MICROSCOPY; X-RAY CRYSTALLOGRAPHY; PHASE PROBLEM; | |
DOI : 10.1107/S2052252518005857 | |
学科分类:数学(综合) | |
来源: International Union of Crystallography | |
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【 摘 要 】
X-ray crystallography and cryo-electron microscopy (cryo-EM) are complementary techniques for structure determination. Crystallography usually reveals more detailed information, while cryo-EM is an extremely useful technique for studying large-sized macromolecules. As the gap between the resolution of crystallography and cryo-EM data narrows, the cryo-EM map of a macromolecule could serve as an initial model to solve the phase problem of crystal diffraction for high-resolution structure determination. FSEARCH is a procedure to utilize the low-resolution molecular shape for crystallographic phasing. The IPCAS (Iterative Protein Crystal structure Automatic Solution) pipeline is an automatic direct-methods-aided dual-space iterative phasing and model-building procedure. When only an electron-density map is available as the starting point, IPCAS is capable of generating a completed model from the phases of the input map automatically, without the requirement of an initial model. In this study, a hybrid method integrating X-ray crystallography with cryo-EM to help with structure determination is presented. With a cryo-EM map as the starting point, the workflow of the method involves three steps. (1) Cryo-EM map replacement: FSEARCH is utilized to find the correct translation and orientation of the cryo-EM map in the crystallographic unit cell and generates the initial low-resolution map. (2) Phase extension: the phases calculated from the correctly placed cryo-EM map are extended to high-resolution X-ray data by non-crystallographic symmetry averaging with phenix.resolve. (3) Model building: IPCAS is used to generate an initial model using the phase-extended map and perform model completion by iteration. Four cases (the lowest cryo-EM map resolution being 6.9 Å) have been tested for the general applicability of the hybrid method, and almost complete models have been generated for all test cases with reasonable Rwork/Rfree. The hybrid method therefore provides an automated tool for X-ray structure determination using a cryo-EM map as the starting point.【 授权许可】
CC BY
【 预 览 】
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RO201910252905147ZK.pdf | 1909KB | ![]() |