Ban N, Freeborn B, Nissen P, Penczek P, Grassucci R A, Sweet R, Frank J, Moore P B, Steitz T A
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06520-8114, USA.
Cell. 1998 Jun 26;93(7):1105-15. doi: 10.1016/s0092-8674(00)81455-5.
The 50S subunit of the ribosome catalyzes the peptidyl-transferase reaction of protein synthesis. We have generated X-ray crystallographic electron density maps of the large ribosomal subunit from Haloarcula marismortui at various resolutions up to 9 A using data from crystals that diffract to 3 A. Positioning a 20 A resolution EM image of these particles in the crystal lattice produced phases accurate enough to locate the bound heavy atoms in three derivatives using difference Fourier maps, thus demonstrating the correctness of the EM model and its placement in the unit cell. At 20 A resolution, the X-ray map is similar to the EM map; however, at 9 A it reveals long, continuous, but branched features whose shape, diameter, and right-handed twist are consistent with segments of double-helical RNA that crisscross the subunit.
核糖体的50S亚基催化蛋白质合成中的肽基转移酶反应。我们利用衍射至3 Å的晶体数据,生成了来自嗜盐嗜碱菌(Haloarcula marismortui)的大核糖体亚基在高达9 Å的各种分辨率下的X射线晶体学电子密度图。将这些颗粒的20 Å分辨率的电子显微镜图像定位在晶格中,产生的相位足够精确,能够使用差分傅里叶图在三种衍生物中定位结合的重原子,从而证明了电子显微镜模型的正确性及其在晶胞中的位置。在20 Å分辨率下,X射线图与电子显微镜图相似;然而,在9 Å时,它揭示了长的、连续但分支的特征,其形状、直径和右手螺旋与贯穿亚基的双螺旋RNA片段一致。