Comparative N-glycan profiling of colorectal cancer cell lines reveals unique bisecting GlcNAc and α-2,3-linked sialic acid determinants are associated with membrane proteins of the more metastatic/aggressive cell lines

Sethi MK, Thaysen-Andersen M, Smith JT, Baker MS, Packer NH, Hancock WS, Fanayan S

PubMed Entry: 24295106

Percentage Abundance
Structure Hex HexNAc Fuc NeuNAc LIM1215 LIM1899 LIM2405 UniCarb-DB
5 3 0 1 1.1 1.8 0.0
5 3 1 1 0.7 1.0 0.5
6 3 0 1 1.3 1.8 1.0
6 4 0 0 0.3 0.0 0.0
6 4 1 0 0.3 0.0 0.0
3 5 0 0 0.3 0.0 0.0
3 5 1 0 0.7 0.0 0.0
3 5 1 0 0.3 0.7 0.0
4 5 0 0 0.5 0.0 0.0
4 5 1 0 0.2 0.3 0.0

    Data Processing

    • Each glycan structure was quantified relative to the total content by integration of the extracted ion chromatogram peak area (EIC).
    • For comparison of glycan abundances the area-under-the-curves (AUCs) of each glycan structure were normalised to the total AUC (for all glycan structures in a sample summed) and expressed as a percentage.

    Additonal Notes

    • Glycoproteins immobilized on PVDF membranes before N-glycans released by PNGase F, isolated and reduced
    • O-glycans are chemically released from the same protein spot by reductive β-elimination
    • Porous graphitised carbon liquid chromatography tandem mass spectrometry
    • Negative ion mode
    • Agilent LCD/MSD Trap XCT Ultra