Highly fucosylated N-glycan ligands for mannan-binding protein expressed specifically on CD26 (DPPVI) isolated from a human colorectal carcinoma cell line, SW1116
Resource
Glycobiology 19 (4): 437-450
Journal
Glycobiology
Pages
437-450
Date Issued
2009
Date
2009
Author(s)
Kawasaki, Nobuko
Lin, Chia-Wei
Inoue, Risa
Khoo, Kay-Hooi
Kawasaki, Nana
Ma, Bruce Yong
Oka, Shogo
Ishiguro, Masaji
Sawada, Toshihiko
Ishida, Hideharu
Hashimoto, Tomohiro
Kawasaki, Toshisuke
Abstract
The 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, multiantennary N-glycans with highly fucosylated polylactosamine-type structures having Leb Lea or tandem repeats of the Lea structure at their nonreducing ends. In this study, we isolated the major MBP-ligand glycoproteins from SW1116 cell lysates with an MBP column and identified them as CD26/dipeptidyl peptidase 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, a 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. ? The Author 2009. Published by Oxford University Press. All rights reserved.
Subjects
CD26; Lea epitope; Mannan-binding lectin; Mannan-binding protein; SW1116
SDGs
Other Subjects
CD98 antigen; dipeptidyl peptidase IV; glycan; glycosidase; ligand; mannan binding lectin; antigen specificity; article; binding affinity; cancer cell culture; colorectal carcinoma; controlled study; enzyme metabolism; epitope mapping; human; human cell; ligand binding; molecular weight; priority journal; protein analysis; protein expression; protein function; protein glycosylation; protein isolation; protein localization; protein structure; tandem repeat; Antigens, CD26; Antigens, CD98 Heavy Chain; Cell Line, Tumor; Colorectal Neoplasms; Epitopes; Fucose; Glycosylation; Humans; Ligands; Mannose-Binding Lectin; Models, Molecular; Neoplasm Proteins; Oligosaccharides; Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; Structure-Activity Relationship