Direct localization of the tRNA--anticodon interaction site on the Escherichia coli 30 S ribosomal subunit by electron microscopy and computerized image averaging.
Direct localization of the tRNA--anticodon interaction site on the Escherichia coli 30 S ribosomal subunit by electron microscopy and computerized image averaging.
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DOI：
10.1016/0022-2836(88)90207-0
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发表时间：
1988-10
影响因子：
5.6
通讯作者：
T. Wagenknecht;J. Frank;M. Boublik;K. Nurse;J. Ofengand
中科院分区：
文献类型：
--
作者：
T. Wagenknecht;J. Frank;M. Boublik;K. Nurse;J. Ofengand
以前的免疫电镜研究表明，缬氨酰-tRNA的反密码子，光交联的核糖体P位点的C1400残基的16 S RNA，位于附近的裂缝的核糖体小亚基ofEscherichia coli。在这项研究中，我们使用单粒子图像平均技术，以证明30 S结合的tRNA分子可以直接定位，而不需要特定的抗体标记。与免疫电镜结果一致，我们发现tRNA分子似乎位于30 S亚基的裂缝深处。我们相信，使用计算机图像平均定位结合到核糖体和其他大分子复合物的配体将成为普遍的，因为上级的灵敏度，精度和客观性，这种技术相比，传统的免疫电子显微镜。
Previous immunoelectron microscopy studies have shown that the anticodon of valyl-tRNA, photocrosslinked to the ribosomal P site at the C1400 residue of the 16 S RNA, is located in the vicinity of the cleft of the small ribosomal subunit ofEscherichia coli. In this study we used single-particle image-averaging techniques to demonstrate that the 30 S-bound tRNA molecule can be localized directly, without the need for specific antibody markers. In agreement with the immunoelectron microscopy results, we find that the tRNA molecule appears to be located deep in the cleft of the 30 S subunit. We believe that the use of computer image averaging to localize ligands bound to ribosomes and other macromolecular complexes will become widespread because of the superior sensitivity, precision and objectivity of this technique compared with conventional immunoelectron microscopy.