Differential expression of ribosomal proteins in a human metastasis model identified by coupling 2-D liquid chromatography and mass spectrometry

Abstract

Background: Proteomic profiling of an experimental tumor metastasis model has the potential to identify gene products that can influence this fatal phenotype of tumor cells. In this study, we focused on the notoriously difficult to assess ribosomal protein component of a pair of cell lines which originate from the same tumor but have opposite metastatic capabilities.

Materials and methods: Cell lysate proteins were separated using a two-dimensional liquid chromatographic system directly coupled to an ESI-TOF mass spectrometer for accurate intact protein MW analysis. Characterization of distinct post-translational modifications and sequence variation within several ribosomal proteins was obtained using monolithic capillary LC/MS/MS, MALDI-MS and -MS/MS.

Results: The combination of these techniques enabled the identification of 45 unique ribosomal proteins, several of which were differentially expressed in metastatic M4A4 cells.

Conclusion: The described proteomic profiling approach enables the identification of phenotype-associated ribosomal proteins for subsequent functional analyses and disease biomarker development.

Figure 1

NPS-RP-HPLC chromatogram of pH fraction 6.0–6.2 with identified ribosomal proteins annotated.

Figure 2

Monolithic LC/MS/MS of tryptic peptide, MNDTVTIR (1–8), from ribosomal protein S24.

Figure 3

MALDI-MS/MS tryptic peptide, AEVQVLVLDGR (1–11), from 60S ribosomal protein L13a.