Department of Bioengineering, Northeastern University, Boston, MA, USA; Barnett Institute for Chemical and Biological Analysis, Northeastern University, Boston, MA, USA.
Department of Biological Sciences, Columbia University, New York, NY, USA.
Trends Biochem Sci. 2019 May;44(5):478-479. doi: 10.1016/j.tibs.2019.01.008. Epub 2019 Feb 18.
Contrary to the textbook model, recent measurements demonstrated unexpected diversity in ribosomal composition that likely enables specialized translational functions. Methods based on liquid chromatography coupled to tandem mass-spectrometry (LC-MS/MS) enable direct quantification of ribosomal proteins with high specificity, accuracy, and throughput. LC-MS/MS can be 'top-down', analyzing intact proteins, or more commonly 'bottom-up', where proteins are digested to peptides prior to analysis. Changes to rRNA can be examined using either LC-MS/MS or sequencing-based approaches. The regulation of protein synthesis by specialized ribosomes can be examined by multiple methods. These include the popular 'Ribo-Seq' method for analyzing ribosome density on a given mRNA, as well as LC-MS/MS approaches incorporating pulse-labelling with stable isotopes (SILAC) to monitor protein synthesis and degradation.
与教科书模型相反，最近的测量结果表明核糖体组成存在出人意料的多样性，这可能使核糖体具有专门的翻译功能。基于液相色谱与串联质谱（LC-MS/MS）的方法能够以高特异性、准确性和高通量直接定量核糖体蛋白。LC-MS/MS 可以是“自上而下”的，分析完整的蛋白质，或者更常见的是“自下而上”的，在这种方法中，蛋白质在分析前被消化成肽。可以使用 LC-MS/MS 或基于测序的方法来检测 rRNA 的变化。通过多种方法可以研究特殊核糖体对蛋白质合成的调控。这些方法包括分析特定 mRNA 上核糖体密度的流行“Ribo-Seq”方法，以及结合稳定同位素脉冲标记（SILAC）的 LC-MS/MS 方法，以监测蛋白质的合成和降解。
