Yeast Ribosomal Protein Populations Characterized by 2D-SEC MS/MS

78 of 79 core yeast ribosomal proteins were identified in the polysome fraction with 93% mean sequence coverage using a two-dimensional size exclusion chromatography tandem mass spectrometry workflow. The method, developed by Andrew J. Spiteri and shared on figshare, delivers four distinct ribosomal subpopulations in under 90 minutes and can resolve 36 of 38 paralogous protein pairs. It also mapped 12 known methylation sites and two phosphorylation sites, enabling site-specific post-translational modification measurements.

Use Cases

• Quantifying isoform-specific incorporation biases based on resolved paralogous protein pairs.
• Measuring site-specific post-translational modifications based on mapped methylation and phosphorylation sites.
• Studying ribosome functional perturbations based on detected changes in polysome profiling ratios under nutrient deprivation or gene knockout conditions.
• Characterizing ribosomal subpopulations based on the separation of polysomes, monosomes, and subunits via two-dimensional chromatography.

Strengths

• Identifies 78 of 79 core yeast ribosomal proteins in the polysome fraction.
• Achieves 93% mean sequence coverage for identified proteins.
• Resolves and differentially quantifies 36 of 38 paralogous protein pairs.
• Maps all 12 known yeast ribosomal methylation sites and two canonical phosphorylation sites.

Limitations

• Column-level documentation is absent; field semantics must be inferred after download.
• Row count is unknown, which may limit suitability assessment.
• Data scale is limited to a 6.1 MB file, suggesting a focused experimental dataset.

Provenance

Source

figshare

Collection Method

Two-dimensional size exclusion chromatography coupled to bottom-up LC-MS/MS.

Freshness

Last updated 2026-05-08 13:08:09; freshness should be verified.