| RRC ID |
53858
|
| Author |
Nakakita S, Itoh A, Nakakita Y, Nonaka Y, Ogawa T, Nakamura T, Nishi N.
|
| Title |
Cooperative Interactions of Oligosaccharide and Peptide Moieties of a Glycopeptide Derived from IgE with Galectin-9.
|
| Journal |
J Biol Chem
|
| Abstract |
We previously showed that galectin-9 suppresses degranulation of mast cells through protein-glycan interaction with IgE. To elucidate the mechanism of the interaction in detail, we focused on identification and structural analysis of IgE glycans responsible for the galectin-9-induced suppression using mouse monoclonal IgE (TIB-141). TIB-141 in combination with the antigen induced degranulation of RBL-2H3 cells, which was almost completely inhibited by human and mouse galectin-9. Sequential digestion of TIB-141 with lysyl endopeptidase and trypsin resulted in the identification of a glycopeptide (H-Lys13-Try3; 48 amino acid residues) with a single N-linked oligosaccharide near the N terminus capable of neutralizing the effect of galectin-9 and another glycopeptide with two N-linked oligosaccharides (H-Lys13-Try1; 16 amino acid residues) having lower activity. Enzymatic elimination of the oligosaccharide chain from H-Lys13-Try3 and H-Lys13-Try1 completely abolished the activity. Removal of the C-terminal 38 amino acid residues of H-Lys13-Try3 with glutamyl endopeptidase, however, also resulted in loss of the activity. We determined the structures of N-linked oligosaccharides of H-Lys13-Try1. The galectin-9-binding fraction of pyridylaminated oligosaccharides contained asialo- and monosialylated bi/tri-antennary complex type oligosaccharides with a core fucose residue. The structures of the oligosaccharides were consistent with the sugar-binding specificity of galectin-9, whereas the nonbinding fraction contained monosialylated and disialylated biantennary complex type oligosaccharides with a core fucose residue. Although the oligosaccharides linked to H-Lys13-Try3 could not be fully characterized, these results indicate the possibility that cooperative binding of oligosaccharide and neighboring polypeptide structures of TIB-141 to galectin-9 affects the overall affinity and specificity of the IgE-lectin interaction.
|
| Volume |
291(2)
|
| Pages |
968-79
|
| Published |
2016-1-8
|
| DOI |
10.1074/jbc.M115.694448
|
| PII |
S0021-9258(20)36227-X
|
| PMID |
26582205
|
| PMC |
PMC4705414
|
| MeSH |
Animals
Antibodies, Monoclonal / metabolism
Cell Degranulation
Cell Line
Chromatography, High Pressure Liquid
Chromatography, Reverse-Phase
Electrophoresis, Polyacrylamide Gel
Enzyme Activation
Galectins / metabolism*
Glycopeptides / isolation & purification*
Glycopeptides / metabolism
Humans
Immunoglobulin E / metabolism*
Mice
Mitogen-Activated Protein Kinases / metabolism
Oligosaccharides / metabolism*
Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / metabolism
Polysaccharides / chemistry
Polysaccharides / metabolism
Protein Binding
Rats
Serine Endopeptidases / metabolism
Serum Albumin, Bovine / metabolism
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Trypsin / metabolism
beta-N-Acetylhexosaminidases / metabolism
|
| IF |
4.238
|
| Times Cited |
6
|
| Resource |
| DNA material |
mIGEH (RDB15983)
mIGEL (RDB15984) |
| Human and Animal Cells |
RBL 2H3(RCB2782) |
