Rose Max, Bobkov Sergey, Ayyer Kartik, Kurta Ruslan P, Dzhigaev Dmitry, Kim Young Yong, Morgan Andrew J, Yoon Chun Hong, Westphal Daniel, Bielecki Johan, Sellberg Jonas A, Williams Garth, Maia Filipe R N C, Yefanov Olexander M, Ilyin Vyacheslav, Mancuso Adrian P, Chapman Henry N, Hogue Brenda G, Aquila Andrew, Barty Anton, Vartanyants Ivan A
Deutsches Elektronen-Synchrotron DESY, Notkestrasse 85, Hamburg D-22607, Germany.
National Research Centre 'Kurchatov Institute', Akademika Kurchatova pl. 1, Moscow 123182, Russia.
IUCrJ. 2018 Sep 18;5(Pt 6):727-736. doi: 10.1107/S205225251801120X. eCollection 2018 Nov 1.
The analysis of a single-particle imaging (SPI) experiment performed at the AMO beamline at LCLS as part of the SPI initiative is presented here. A workflow for the three-dimensional virus reconstruction of the PR772 bacteriophage from measured single-particle data is developed. It consists of several well defined steps including single-hit diffraction data classification, refined filtering of the classified data, reconstruction of three-dimensional scattered intensity from the experimental diffraction patterns by orientation determination and a final three-dimensional reconstruction of the virus electron density without symmetry constraints. The analysis developed here revealed and quantified nanoscale features of the PR772 virus measured in this experiment, with the obtained resolution better than 10 nm, with a clear indication that the structure was compressed in one direction and, as such, deviates from ideal icosahedral symmetry.
本文介绍了作为单粒子成像（SPI）计划一部分，在直线加速器相干光源（LCLS）的AMO光束线进行的单粒子成像（SPI）实验分析。开发了一种从测量的单粒子数据重建PR772噬菌体三维病毒结构的工作流程。它包括几个明确的步骤，包括单次衍射数据分类、对分类数据进行精细滤波、通过取向确定从实验衍射图案重建三维散射强度，以及在无对称性约束下对病毒电子密度进行最终的三维重建。此处开展的分析揭示并量化了在该实验中测量的PR772病毒的纳米级特征，获得的分辨率优于10 nm，清楚表明该结构在一个方向上被压缩，因此偏离了理想的二十面体对称性。
