Protein synthesis is a complex, multi-step process that involves large conformational changes of the ribosome and protein factors of translation. Over the last decade, Förster resonance energy transfer (FRET) has become instrumental for studying structural rearrangements of the translational apparatus. Here, we discuss the design of ensemble and single-molecule (sm) FRET assays of translation. We describe a number of experimental strategies that can be used to introduce fluorophores into the ribosome, tRNA, mRNA and protein factors of translation. Alternative approaches to tethering of translation components to the microscope slide in smFRET experiments are also reviewed. Finally, we discuss possible challenges in the interpretation of FRET data and ways to address these challenges.

中文翻译：

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蛋白质合成的整体和单分子 FRET 研究

蛋白质合成是一个复杂的多步骤过程，涉及核糖体和蛋白质翻译因子的巨大构象变化。在过去的十年中，福斯特共振能量转移 (FRET) 已成为研究平移装置结构重排的工具。在这里，我们讨论翻译的整体和单分子 (sm) FRET 检测的设计。我们描述了一些可用于将荧光团引入核糖体、tRNA、mRNA 和翻译蛋白因子的实验策略。还审查了在 smFRET 实验中将翻译组件连接到显微镜载玻片的替代方法。最后，我们讨论了 FRET 数据解释中可能面临的挑战以及解决这些挑战的方法。