Title : N-glycan structures and N-glycosylation
sites of mouse soluble
intercellular adhesion molecule-1 revealed by MALDI-TOF and FTICR mass spectrometry
Abstract :
- Intercellular adhesion molecule-1 ( ICAM-1 ) is a heavily N-glycosylated transmembrane protein comprising five extracellular Ig-like domains
- The soluble isoform of ICAM-1 isoform of ICAM-1 ( sICAM-1 ), consisting of its extracellular part, is elevated in the cerebrospinal fluid of patients with severe brain trauma
- In mouse astrocytes, recombinant mouse sICAM-1 induces the production of the CXC chemokine macrophage inflammatory protein-2 ( MIP-2 )
- MIP-2 induction is glycosylation dependent, as it is strongly enhanced when sICAM-1 carries sialylated, complex-type N-glycans as synthesized by wild-type Chinese hamster ovary (CHO) cells
- The present study was aimed at elucidating the N-glycosylation of mouse sICAM-1 expressed in wild-type CHO cells with regard to sialylation, N-glycan profile, and N-glycosylation sites
- Ion-exchange chromatography and matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) of the released N-glycans showed that sICAM-1 mostly carried di- and trisialylated complex-type N-glycans with or without one fucose
- In some sialylated N-glycans, one N-acetylneuraminic acid was replaced by N-glycolylneuraminic acid, and approximately 4% carried a higher number of sialic acid residues than of antennae
- The N-glycosylation sites of mouse sICAM-1 were analyzed by MALDI-Fourier transform ion cyclotron resonance (FTICR)-MS and nanoLC- ESI-FTICR-MS of tryptic digests of mouse sICAM-1 expressed in the Lec1 mutant of CHO cells
- All nine consensus sequences for N-glycosylation were found to be glycosylated
- These results show that the N-glycans that enhance the MIP-2-inducing activity of mouse sICAM-1 are mostly di- and trisialylated complex-type N-glycans including a small fraction carrying more sialic acid residues than antennae and that the nine N-glycosylation sites of mouse sICAM-1 are all glycosylated
Output (sent_index, trigger,
protein,
sugar,
site):
- 0. N-glycosylation, , intercellular adhesion molecule-1, -, sites
- 0. molecule-1, , intercellular adhesion molecule-1, N-glycan structures, -
- 1. N-glycosylated, , Intercellular adhesion molecule-1, -, -
- 1. N-glycosylated, , protein, -, -
- 10. N-glycosylation, , sICAM-1, -, sites
- 10. glycosylated, , sICAM-1, -, sites
- 10. trisialylated, , -, a small fraction, -
- 10. trisialylated, , -, mostly di- and trisialylated complex-type N-glycans, -
- 10. trisialylated, , -, the N-glycans, -
- 4. carries, , sICAM-1, complex-type N-glycans, -
- 4. sialylated, , -, complex-type N-glycans, -
- 5. N-glycosylation, , -, -, sites
- 5. N-glycosylation, , sICAM-1, -, -
- 6. carried, , sICAM-1, di-, -
- 6. carried, , sICAM-1, trisialylated complex-type N-glycans, -
- 6. trisialylated, , -, trisialylated complex-type N-glycans, -
- 7. sialylated, , -, some sialylated N-glycans, -
- 8. N-glycosylation, , sICAM-1, -, sites
- 9. glycosylated, , -, -, -
Output(Part-Of) (sent_index,
protein,
site):
- 0. intercellular adhesion molecule-1, sites
- 10. sICAM-1, sites
- 8. sICAM-1, sites
*Output_Site_Fusion* (sent_index,
protein,
sugar,
site):