Identification of proteins bearing beta1-6 branched N-glycans in human melanoma cell lines from different progression stages by tandem mass spectrometry analysis.
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.
Przybyło
Małgorzata
M
Department of Glycoconjugate Biochemistry, Institute of Zoology, Jagiellonian University, Ingardena 6, 30-060 Kraków, Poland.
Martuszewska
Danuta
D
Pocheć
Ewa
E
Hoja-Łukowicz
Dorota
D
Lityńska
Anna
A
D045744:Cell Line, Tumor; D006801:Humans; D008545:Melanoma / Q000737:chemistry*; D025942:Oligosaccharides, Branched-Chain / Q000737:chemistry; D010835:Phytohemagglutinins / Q000737:chemistry; D017026:Swainsonine / Q000494:pharmacology; D053719:Tandem Mass Spectrometry; D014945:Wound Healing / Q000187:drug effects
D025942:Oligosaccharides, Branched-Chain; D010835:Phytohemagglutinins; C100156:leukoagglutinins, plants; D017026:Swainsonine