| N-glycan structures and N-glycosylation sites of mouse soluble intercellular adhesion molecule-1 revealed by MALDI-TOF and FTICR mass spectrometry. | |
MedLine Citation:
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PMID: 16877748 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
Authors:
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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:
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Type: Journal Article Date: 2006-07-28 |
Journal Detail:
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Title: Glycobiology Volume: 16 ISSN: 0959-6658 ISO Abbreviation: Glycobiology Publication Date: 2006 Nov |
Date Detail:
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Created Date: 2006-10-02 Completed Date: 2007-03-16 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9104124 Medline TA: Glycobiology Country: England |
Other Details:
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Languages: eng Pagination: 1033-44 Citation Subset: IM |
Affiliation:
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Institute of Pharmaceutical Sciences, Department of Chemistry and Applied Biosciences, ETH Zurich, Switzerland. vivianne.otto@pharma.ethz.ch |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals CHO Cells Consensus Sequence Cricetinae Cricetulus Glycosylation Intercellular Adhesion Molecule-1 / genetics, metabolism* Mass Spectrometry Mice Mutation N-Acetylneuraminic Acid / metabolism Polysaccharides / metabolism* Protein Isoforms / chemistry Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization |
| Chemical | |
Reg. No./Substance:
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0/Polysaccharides; 0/Protein Isoforms; 126547-89-5/Intercellular Adhesion Molecule-1; 131-48-6/N-Acetylneuraminic Acid |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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