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Approaches for Studying Ribosome Specialization.

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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… Click to show full abstract

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.

Keywords: studying ribosome; ribosome specialization; approaches studying

Journal Title: Trends in biochemical sciences
Year Published: 2019

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