Glycobiology Advance Access published online on January 7, 2009
Glycobiology, doi:10.1093/glycob/cwn158
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Highly Fucosylated N-Glycan Ligands for Mannan-binding Protein Expressed Specifically on CD26 (DPPVI) Isolated from a Human Colorectal Carcinoma Cell Line, SW1116
2 Research Center for Glycobiotechnology, Ritsumeikan University, Shiga 525-8577, Japan
3 Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan
4 Graduate Institute of Biochemical Sciences, National Taiwan University, Taipei 106, Taiwan
5 Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, Tokyo 158-8501, Japan
6 Human Health Sciences, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan
7 Suntory Institute for Bioorganic Research, Osaka 618-8503, Japan
8 Faculty of Applied Biological Sciences, Gifu University, Gifu 501-1193, Japan
9 Faculty of Regional Studies, Gifu University, Gifu 501-1193, Japan
1 To whom corresponence should be addressed: Tel: +81-77-3444; Fax: +81-77-3496; e-mail: tkawasak{at}fc.ritsumei.ac.jp
Received on September 30, 2008; accepted on December 27, 2008
Serum mannan-binding protein (MBP) is a host defense C-type lectin specific for mannose, N-acetylglucosamine and fucose residues, and exhibits growth inhibitory activity toward human colorectal carcinoma cells. The MBP-ligand oligosaccharides (MLO) isolated from a human colorectal carcinoma cell line, SW1116, are large, multi-antennary N-glycans with highly fucosylated polylactosamine-type structures having Leb-Lea or tandem repeats of the Lea structure at their non-reducing ends. In this study, we isolated the major MBP-ligand glycoproteins (MLGPs) from SW1116 cell lysates with an MBP column and identified them as CD26/dipeptidylpeptidase IV (DPPIV) (110 kDa) and CD98 heavy chain (CD98hc)/4F2hc (82 kDa Glycosidase digestion revealed that CD26 contained such complex-type N-glycans that appear to mediate the MBP binding. MALDI-MS of the N-glycans released from CD26 by PNGase F demonstrated conclusively that CD26 is the major MLO-carrying protein. More interestingly, comparison of the N-glycans released from the MBP-binding and non-MBP-binding glycopeptides suggested that complex type N-glycans carrying a minimum of 4 Lea /Leb epitopes arranged either as multimeric tandem repeats or terminal epitopes on multiantennary structures are critically important for the high affinity binding to MBP. Analysis of the N-glycan attachment sites demonstrated that the high affinity MLO was expressed preferentially at some N-glycosylation sites, but this site preference was not so stringent. Finally, hypothetical 3D models of tandem repeats of the Lea epitope and the MBP-Lewis oligosaccharide complex were presented.
Key words: CD26 / Mannan-binding lectin / Mannan-binding protein / Lea epitope / SW1116
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