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Trends Biochem Sci. 2019 May;44(5):478-479. doi: 10.1016/j.tibs.2019.01.008. Epub 2019 Feb 18.

Approaches for Studying Ribosome Specialization.

Author information

1
Department of Bioengineering, Northeastern University, Boston, MA, USA; Barnett Institute for Chemical and Biological Analysis, Northeastern University, Boston, MA, USA.
2
Department of Biological Sciences, Columbia University, New York, NY, USA.
3
Department of Bioengineering, Northeastern University, Boston, MA, USA; Barnett Institute for Chemical and Biological Analysis, Northeastern University, Boston, MA, USA; Department of Biology, Northeastern University, Boston, MA, USA. Electronic address: nslavov@alum.mit.edu.

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.

PMID:
30792028
DOI:
10.1016/j.tibs.2019.01.008

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