Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California 91125, United States.
J Am Chem Soc. 2011 Sep 28;133(38):14936-9. doi: 10.1021/ja206626g. Epub 2011 Sep 2.
As newly synthesized proteins emerge from the ribosome, they interact with a variety of cotranslational cellular machineries that facilitate their proper folding, maturation, and localization. These interactions are essential for proper function of the cell, and the ability to study these events is crucial to understanding cellular protein biogenesis. To this end, we have developed a highly efficient method to generate ribosome-nascent chain complexes (RNCs) site-specifically labeled with a fluorescent dye on the nascent polypeptide. The fluorescent RNC provides real-time, quantitative information on its cotranslational interaction with the signal recognition particle and will be a valuable tool in elucidating the role of the translating ribosome in numerous biochemical pathways.
当新合成的蛋白质从核糖体中出现时,它们会与各种共翻译细胞机制相互作用,从而促进其正确折叠、成熟和定位。这些相互作用对于细胞的正常功能至关重要,而研究这些事件的能力对于理解细胞蛋白质生物发生至关重要。为此,我们开发了一种非常有效的方法,可以在新生多肽上对核糖体-新生链复合物(RNC)进行特异性荧光标记。荧光 RNC 提供了关于其与信号识别颗粒共翻译相互作用的实时、定量信息,它将成为阐明翻译核糖体在众多生化途径中的作用的有价值工具。