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 2 0 0 1.4 1.6 1.1
6 2 0 0 6.2 5.1 5.7
7 2 0 0 7.8 6.3 7.8
7 2 0 0 2.3 2.3 2.6
8 2 0 0 23.7 19.4 21.7
9 2 0 0 45.4 46.0 32.5
10 2 0 0 1.6 1.7 0.5
6 3 0 0 0.4 0.0 0.0
5 4 0 0 0.4 0.0 0.0
5 4 0 0 0.5 0.0 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