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Identification of proteins bearing beta1-6 branched N-glycans in human melanoma cell lines from different progression stages by tandem mass spectrometry analysis.

Abstract
The common structural alterations in the cell-surface glycoproteins concern the highly elevated expression of tri- and tetra-antennary beta1-6-N-acetylglucosamine (beta1-6 GlcNAc) bearing N-glycans, which are recognised by Phaseolus vulgaris agglutinin (PHA-L). In this report we identified proteins bearing beta1-6 GlcNAc branched N-glycans in three human melanoma cell lines: WM35--from the primary tumour site, as well as WM239 and WM9 from different metastatic sites: the skin and the lymph node, respectively, by tandem mass spectrometry (MS/MS) on PHA-L agarose bound material, followed by immunochemical identification. Our results show that melanoma cell lines differ from each other in the number of N-glycoproteins bearing beta1-6 GlcNAc branched oligosaccharides. Among identified proteins the largest group consists of integrin subunits. In addition, L1-CAM, Mac-2 binding protein, melanoma cell adhesion molecule, intercellular adhesion molecule, melanoma associated antigen, tumour rejection antigen-1, melanoma-associated chondroitin sulfate proteoglycan 4 and lysosome-associated membrane protein (LAMP-1) were found. It was indicated that WM35 cell line showed the lowest number of proteins possessing beta1-6 GlcNAc branched N-glycans in comparison to metastatic WM9 and WM239 cell lines. Our data suggest that changes in the number of proteins being a substrate for GlcNAc-TV are better correlated with melanoma development and progression than with expression of cell adhesion molecules.
AuthorsMałgorzata Przybyło, Danuta Martuszewska, Ewa Pocheć, Dorota Hoja-Łukowicz, Anna Lityńska
JournalBiochimica et biophysica acta (Biochim Biophys Acta) Vol. 1770 Issue 9 Pg. 1427-35 (Sep 2007) ISSN: 0006-3002 [Print] Netherlands
PMID17600626 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Oligosaccharides, Branched-Chain
  • Phytohemagglutinins
  • leukoagglutinins, plants
  • Swainsonine
Topics
  • Cell Line, Tumor
  • Humans
  • Melanoma (chemistry)
  • Oligosaccharides, Branched-Chain (chemistry)
  • Phytohemagglutinins (chemistry)
  • Swainsonine (pharmacology)
  • Tandem Mass Spectrometry
  • Wound Healing (drug effects)

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