Walz T, Jamieson S J, Bowers C M, Bullough P A, Hunter C N
Department of Molecular Biology and Biotechnology, University of Sheffield, Firth Court Western Bank, Sheffield, S10 2TN, UK.
J Mol Biol. 1998 Oct 2;282(4):833-45. doi: 10.1006/jmbi.1998.2050.
Three photosynthetic complexes, light-harvesting complex 2 (LH2), light-harvesting complex 1 (LH1), and the reaction centre-light-harvesting complex 1 photounit (RC-LH1), were purified from a single species of a purple bacterium, Rhodobacter sphaeroides, and reconstituted into two-dimensional (2-D) crystals. Vesicular 2-D crystals of LH1 and RC-LH1 were imaged in negative stain and projection maps at 25 A resolution were produced. The rings formed by LH1 have approximately the same mean diameter as the LH1 rings from Rhodospirillum rubrum ( approximately 90 A) and therefore are likely to be composed of 15 to 17 alphabeta subunits. In the projection map calculated from the RC-LH1 2-D crystals, the reaction centre is represented by an additional density in the centre of the ring formed by the LH1 subunits. The marked improvement of shape and fine structure after a rotational pre-alignment of the RC-LH1 unit cells before averaging strongly suggests that the RC is not in a unique orientation within the LH1 rings. Tubular crystals of LH2 showed a high degree of order and allowed calculation of a projection map at 6 A resolution from glucose-embedded specimens. The projection structure shows a ring of nine alphabeta subunits. Variation of the alpha-helical projection densities suggests that the 9-fold symmetry axis is tilted with respect to the membrane normal.
从一种紫色细菌球形红杆菌中纯化出三种光合复合物,即捕光复合物2(LH2)、捕光复合物1(LH1)和反应中心 - 捕光复合物1光单元(RC-LH1),并将它们重构为二维(2-D)晶体。对LH1和RC-LH1的囊泡状二维晶体进行负染色成像,并生成了分辨率为25埃的投影图。由LH1形成的环的平均直径与来自红螺菌的LH1环的平均直径大致相同(约90埃),因此可能由15至17个αβ亚基组成。在根据RC-LH1二维晶体计算出的投影图中,反应中心由LH1亚基形成的环中心的额外密度表示。在平均之前对RC-LH1晶胞进行旋转预对齐后,形状和精细结构有明显改善,这强烈表明RC在LH1环内并非处于唯一取向。LH2的管状晶体显示出高度有序性,并能够从葡萄糖包埋的标本中计算出分辨率为6埃的投影图。投影结构显示有一个由九个αβ亚基组成的环。α螺旋投影密度的变化表明9重对称轴相对于膜法线倾斜。