Ludtke Steven J, Chen Dong-Hua, Song Jiu-Li, Chuang David T, Chiu Wah
National Center for Macromolecular Imaging, Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030 USA.
Structure. 2004 Jul;12(7):1129-36. doi: 10.1016/j.str.2004.05.006.
We present a reconstruction of native GroEL by electron cryomicroscopy (cryo-EM) and single particle analysis at 6 A resolution. alpha helices are clearly visible and beta sheet density is also visible at this resolution. While the overall conformation of this structure is quite consistent with the published X-ray data, a measurable shift in the positions of three alpha helices in the intermediate domain is observed, not consistent with any of the 7 monomeric structures in the Protein Data Bank model (1OEL). In addition, there is evidence for slight rearrangement or flexibility in parts of the apical domain. The 6 A resolution cryo-EM GroEL structure clearly demonstrates the veracity and expanding scope of cryo-EM and the single particle reconstruction technique for macromolecular machines.
我们通过电子冷冻显微镜(cryo-EM)和单颗粒分析,以6埃分辨率呈现了天然GroEL的重建结构。α螺旋清晰可见,在此分辨率下β折叠密度也可见。虽然该结构的整体构象与已发表的X射线数据相当一致,但观察到中间结构域中三个α螺旋的位置有可测量的位移,这与蛋白质数据库模型(1OEL)中的7个单体结构均不一致。此外,有证据表明顶端结构域的部分存在轻微重排或灵活性。6埃分辨率的cryo-EM GroEL结构清楚地证明了cryo-EM和用于大分子机器的单颗粒重建技术的准确性和不断扩大的应用范围。