Document Detail

N-glycan structures and N-glycosylation sites of mouse soluble intercellular adhesion molecule-1 revealed by MALDI-TOF and FTICR mass spectrometry.
MedLine Citation:
PMID:  16877748     Owner:  NLM     Status:  MEDLINE    
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 (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.
Vivianne I Otto; Eugen Damoc; Leah N Cueni; Thomas Schürpf; Renate Frei; Sarah Ali; Nico Callewaert; Adrian Moise; Julie A Leary; Gerd Folkers; Michael Przybylski
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Publication Detail:
Type:  Journal Article     Date:  2006-07-28
Journal Detail:
Title:  Glycobiology     Volume:  16     ISSN:  0959-6658     ISO Abbreviation:  Glycobiology     Publication Date:  2006 Nov 
Date Detail:
Created Date:  2006-10-02     Completed Date:  2007-03-16     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9104124     Medline TA:  Glycobiology     Country:  England    
Other Details:
Languages:  eng     Pagination:  1033-44     Citation Subset:  IM    
Institute of Pharmaceutical Sciences, Department of Chemistry and Applied Biosciences, ETH Zurich, Switzerland.
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MeSH Terms
CHO Cells
Consensus Sequence
Intercellular Adhesion Molecule-1 / genetics,  metabolism*
Mass Spectrometry
N-Acetylneuraminic Acid / metabolism
Polysaccharides / metabolism*
Protein Isoforms / chemistry
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Reg. No./Substance:
0/Polysaccharides; 0/Protein Isoforms; 126547-89-5/Intercellular Adhesion Molecule-1; 131-48-6/N-Acetylneuraminic Acid

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