Klostermeier Dagmar, Sears Pamela, Wong Chi-Huey, Millar David P, Williamson James R
The Scripps Research Institute, Department of Molecular Biology, La Jolla, CA 92037, USA.
Nucleic Acids Res. 2004 May 17;32(9):2707-15. doi: 10.1093/nar/gkh588. Print 2004.
In one of the first steps of prokaryotic ribosome assembly, the ribosomal protein S15 binds to a three-way junction in the central domain of the 16S rRNA. Binding causes a conformational change that is required for subsequent binding events. Using a novel fluorescence resonance energy transfer assay with three fluorophores, two on the RNA and one on the S15 protein, small-molecule libraries can be screened for potential inhibitors of this initial step in ribosome assembly. The employment of three fluorophores allows both the conformational change of the RNA and the binding of S15 to be monitored in a single assay.
在原核生物核糖体组装的最初步骤之一中,核糖体蛋白S15与16S rRNA中央结构域的一个三向接头结合。这种结合会引起构象变化,而后续的结合事件需要这种构象变化。利用一种新型的荧光共振能量转移测定法,该方法使用三个荧光团,其中两个位于RNA上,一个位于S15蛋白上,可以筛选小分子文库,以寻找核糖体组装这一初始步骤的潜在抑制剂。使用三个荧光团能够在一次测定中同时监测RNA的构象变化以及S15的结合情况。