ジャーナル: Nat Commun / 年: 2014 タイトル: An atomic model of brome mosaic virus using direct electron detection and real-space optimization. 著者: Zhao Wang / Corey F Hryc / Benjamin Bammes / Pavel V Afonine / Joanita Jakana / Dong-Hua Chen / Xiangan Liu / Matthew L Baker / Cheng Kao / Steven J Ludtke / Michael F Schmid / Paul D Adams / Wah Chiu / 要旨: Advances in electron cryo-microscopy have enabled structure determination of macromolecules at near-atomic resolution. However, structure determination, even using de novo methods, remains ...Advances in electron cryo-microscopy have enabled structure determination of macromolecules at near-atomic resolution. However, structure determination, even using de novo methods, remains susceptible to model bias and overfitting. Here we describe a complete workflow for data acquisition, image processing, all-atom modelling and validation of brome mosaic virus, an RNA virus. Data were collected with a direct electron detector in integrating mode and an exposure beyond the traditional radiation damage limit. The final density map has a resolution of 3.8 Å as assessed by two independent data sets and maps. We used the map to derive an all-atom model with a newly implemented real-space optimization protocol. The validity of the model was verified by its match with the density map and a previous model from X-ray crystallography, as well as the internal consistency of models from independent maps. This study demonstrates a practical approach to obtain a rigorously validated atomic resolution electron cryo-microscopy structure.
EMPIAR-10010 (タイトル: Full virus map of Brome Mosaic Virus (micrographs and particle coordinates) Data size: 1.7 TB Data #1: Brome Mosaic Virus micrographs - non gain corrected [micrographs - multiframe] Data #2: Brome Mosaic Virus micrographs - gain corrected [micrographs - multiframe]) EMPIAR-10011 (タイトル: Full virus map of Brome Mosaic Virus (picked particles) Data size: 23.1 Data #1: Brome Mosaic Virus boxed particles [picked particles - single frame - processed])