PMID: 12545205

 

    Legend: Sugar

Title : Carbohydrate moieties of N-cadherin from human melanoma cell lines

Abstract :

1. Expression of N-cadherin an adhesion molecule of the cadherin family, in tumor cells is associated with their increased invasive potential
2. Many studies suggested the role of N-linked oligosaccharides as important factors that contribute to metastasis by influencing tumor cell invasion and adhesion
3. N-cadherin is a heavily glycosylated protein
4. We have analysed the carbohydrate profile of this protein synthesized in human melanoma cell lines: WM35 from the primary tumor site and WM239, WM9, and A375 from different metastatic sites
5. N-cadherin was immunoprecipitated with anti-human N-cadherin polyclonal antibodies
6. Characterisation of its carbohydrate moieties was carried out by SDS /PAGE electrophoresis and blotting, followed by immunochemical identification of the N-cadherin polypeptides and analysis of their glycans using highly specific digoxigenin or biotin labelled lectins
7. The positive reaction of N-cadherin from the WM35 cell line with Galanthus nivalis agglutinin ( GNA ), Datura stramonium agglutinin ( DSA ) and Sambucus nigra agglutinin ( SNA ) indicated the presence of high-mannose type glycans and biantennary complex type oligosaccharides with alpha2-6 linked sialic acid.
8. N-cadherin from WM239, WM9, and A375 cell lines gave a positive reaction with Phaseolus vulgaris leukoagglutinin ( L-PHA ) and lotus Tetragonolobus purpureas agglutinin ( LTA )
9. This indicated the presence of tri- or tetra-antennary complex type glycans with alpha-fucose.
10. In addition, N-cadherin from WM9 (lymphomodus metastatic site ) and A375 (solid tumor metastatic site ) contained complex type chains with alpha2-3 sialic acid (positive reaction with Maackia amurensis agglutinin--MAA)
11. The results demonstrated that N-glycans of N-cadherin are altered in metastatic melanomas in a way characteristic for invasive tumor cells