SCHIMMEL PRT-CRYSTAL STRUCTURE OF TRBP111/TRNA COMPLEX

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 Trbp111 是一种古老的结构特异性 tRNA 识别结构域，存在于许多新型人类细胞凋亡相关细胞因子、氨酰基-tRNA 合成酶和蛋白质合成装置的其他组件中。该结构域的功能是识别 tRNA 的一般 L 形状（无论序列如何），并将其递送至其受体酶以进行有效的氨酰化。我想在原子水平上了解这种结构特异性 tRNA 识别的本质。因此，我确定了细菌 Trbp111 的晶体结构，它揭示了具有两个 OB 折叠结构域的同型二聚体和一个新的二聚体界面（Swairjo 等人，EMBO J，19：6287，2000）。我现在正在研究 tRNA 复合物的晶体结构。我已经生产了 Aquifex aeolicus Trbp111 与大肠杆菌 tRNAfMet 复合的晶体，在内部设备上衍射至 7.0 ¿我希望在 SSRL 收集更高分辨率的 X 射线数据，以便确定复杂的结构。该项目是我们研究最近在基因组数据库中发现的几个新颖的非特异性 RNA 识别域的总体兴趣的一部分（Avarind 和 Koonin, J. Mol. Evol., 38: 291, 1999）。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Trbp111 is an ancient, structure-specific tRNA recognition domain found in many novel, human apoptosis-related cytokines, aminoacyl-tRNA synthetases, and other components of the protein synthesis apparatus. The function of this domain is to recognize the general L-shape of tRNA, regardless of sequence, and deliver it to its receptor enzyme for efficient aminoacylation. I want to understand the nature of this structure-specific tRNA recognition at an atomic level. Therefore, I determined the crystal structure of bacterial Trbp111 which revealed a homodimer with two OB-fold domains and a novel dimer interface (Swairjo et al., EMBO J, 19: 6287, 2000). I am now pursuing the crystal structure of the complex with tRNA. I have produced crystals of Aquifex aeolicus Trbp111 complexed with E. coli tRNAfMet which diffract to 7.0 ¿ on in-house equipment. I hope to collect higher-resolution X-ray data at SSRL that would allow for determination of the complex structure. This project is part of our general interest in studying several, novel, non-specific RNA recognition domains identified recently in genomic databases (Avarind and Koonin, J. Mol. Evol., 38: 291, 1999).