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Author:
Date:
2015-08-20
[Abstract] Telomerase is a large ribonucleoprotein complex that replicates the linear chromosome ends in most eukaryotes. Large-scale preparation of the telomerase core components in vitro has long been a big challenge in this field, hindering the understanding of the catalytic mechanism of telomerase, as well as slowing down the development of telomerase inhibitors for cancer therapy. We have successfully developed a protocol for large-scale preparation of the TRBD-CR4/5 complex of the medaka telomerase in vitro, and used this method to study the high-resolution structure of the TRBD-CR4/5 complex by X-ray crystallography. This procedure may be also adapted to purify other protein-RNA complexes for structural studies.
[摘要] 端粒酶是在大多数真核生物中复制线性染色体末端的大核糖核蛋白复合物。 体外大规模制备端粒酶核心组分长期以来一直是该领域的一个大挑战，阻碍了对端粒酶的催化机理的理解，以及减慢端粒酶抑制剂对癌症的发展 治疗。 我们成功地开发了用于大规模制备体外的medaka端粒酶的TRBD-CR4/5复合物的方案，并且使用该方法研究TRBD-CR4/5复合物的高分辨率结构， 5复合物通过X射线晶体学。 该程序也可以适于纯化其它蛋白质-RNA复合物用于结构研究。
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