Skip to main content
. Author manuscript; available in PMC: 2020 Feb 1.
Published in final edited form as: Trends Biochem Sci. 2018 Nov 22;44(2):95–109. doi: 10.1016/j.tibs.2018.10.009

Figure 4. Identifying altered RP stoichiometry and PTMs.

Figure 4.

A) For decades, the gold standard approach for isolating RPs in the context of intact, functional, ribosomes has been sucrose gradient centrifugation. B) Affinity purification is a powerful means of identifying differential RP association with complexes, however it cannot definitively say whether an RP is resident in a ribosome or represents an extra-ribosomal population of the RP. C) A combined approach whereby affinity purification is performed on sucrose gradient fractions allows the advantages of affinity purification to be applied to samples where the RP is known to be ribosome-resident. D) Heterogeneity amongst ribosomal protein modifications is a promising new area of research, and the methods required to explore this are an extension of those for identifying changes to RP association. Protease-digested peptides from sucrose gradient fractions or affinity purification can be enriched for a particular PTM of choice, either individually, or serially whereby the flow-through of one enrichment is applied to the next enrichment process.