Trachtenberg S, DeRosier D J, Zemlin F, Beckmann E
Department of Membrane and Ultrastructure Research, The Hebrew University-Hadassah Medical School, Jerusalem 91120, Israel.
J Mol Biol. 1998 Mar 6;276(4):759-73. doi: 10.1006/jmbi.1997.1537.
Using a liquid-helium-cooled superconducting electron cryo-microscope, we obtained low-dose images of negatively stained preparations at 4 K and collected structural data to 1/9.6 -1 for flagellar filaments from the strain SJW117 of Salmonella typhimurium (serotype gt). The subunits of this left-handed, straight filament are non-helically perturbed in a pairwise manner. The perturbation corresponds to an alternating conformation in every other row of subunits. These are the 5-start rows and, necessarily, the resulting structure has a seam. The perturbation is not confined to the outside but extends into the structure. We separated the non-symmetric and symmetric parts of the structural data and generated a three-dimensional reconstruction from the latter. The resulting density map is a structure similar in domain organization to the left-handed filament of S. typhimurium SJW1660. Filtered images generated from the non-symmetric component show an ordered and polar structure. The nature of the perturbation was analyzed by model building using a sphere to represent the subunit at low resolution. A lateral shift of approximately 10 degrees mimics the perturbation.
我们使用液氦冷却的超导电子冷冻显微镜,在4 K下获得了鼠伤寒沙门氏菌(血清型gt)SJW117菌株鞭毛丝的负染色制剂的低剂量图像,并收集了分辨率达到1/9.6 Å的结构数据。这种左手性直丝的亚基以成对方式受到非螺旋性扰动。这种扰动对应于每隔一排亚基的交替构象。这些是五起始排,因此,所得结构有一条缝。这种扰动不仅局限于外部,还延伸到结构内部。我们分离了结构数据的非对称部分和对称部分,并从后者生成了三维重建。所得的密度图是一个在结构域组织上与鼠伤寒沙门氏菌SJW1660的左手性丝相似的结构。从非对称成分生成的滤波图像显示出有序的极性结构。通过使用球体在低分辨率下代表亚基进行模型构建,分析了扰动的性质。大约10度的横向位移模拟了这种扰动。