Department of Chemistry, University of Pennsylvania, Philadelphia, PA 19104-6323, USA.
Bioconjug Chem. 2013 Jul 17;24(7):1186-90. doi: 10.1021/bc400062d. Epub 2013 Jun 17.
When suitably labeled bulk tRNAs are transfected into cells they give rise to FRET (fluorescence resonance energy transfer) signals via binding to ribosomes that provide a measure of total protein synthesis. Application of this approach to monitoring rates of specific protein synthesis requires achieving a very high signal-to-noise ratio. Such high ratios may be attainable using LRET (luminescence resonance energy transfer) in place of FRET. Lanthanide complexes containing an antenna chromophore are excellent LRET donors. Here we describe the synthesis of a Phe-tRNA(Phe) labeled with a Tb(3+) complex, denoted Tb(3+)-Phe-tRNA(Phe) that, notwithstanding the bulkiness of the Tb(3+) complex, is active in protein synthesis.
当合适标记的大量 tRNA 转染到细胞中时，它们会通过与核糖体结合产生 FRET（荧光共振能量转移）信号，该信号可作为总蛋白质合成的度量。通过这种方法监测特定蛋白质合成速率需要获得非常高的信噪比。通过使用 LRET（发光共振能量转移）代替 FRET，可能可以达到这样的高比值。含有天线生色团的镧系元素配合物是极好的 LRET 供体。在这里，我们描述了用 Tb(3+) 配合物标记的 Phe-tRNA(Phe)的合成，记为 Tb(3+)-Phe-tRNA(Phe)，尽管 Tb(3+) 配合物很大，但它在蛋白质合成中是有活性的。
