Concepts and principles of glycobiology

Opdenakker, Ghislain; Rudd, Pauline M.; Ponting, Christopher; Dwek, Raymond A.

doi:10.1096/fasebj.7.14.8224606

Cited by 201 publications

(145 citation statements)

References 5 publications

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“…N-Linked glycosylation, which adds oligosaccharides to the nitrogen in the side-chain amide of asparagine residues, requires the asparagines in a consensus sequence of Asn-X-Ser/ Thr (where X is any amino acid except proline) (22,23). However, not all consensus sites are actually glycosylated because the oligosaccharide might be trimmed and elaborated during transit through the endoplasmic reticulum and the Golgi apparatus (23).…”

Section: Discussionmentioning

confidence: 99%

N-Linked Glycosylation Regulates Human Proteinase-activated Receptor-1 Cell Surface Expression and Disarming via Neutrophil Proteinases and Thermolysin

Xiao

Morice

Compton

et al. 2011

Journal of Biological Chemistry

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Section: Discussionmentioning

confidence: 99%

N-Linked Glycosylation Regulates Human Proteinase-activated Receptor-1 Cell Surface Expression and Disarming via Neutrophil Proteinases and Thermolysin

Xiao

Morice

Compton

et al. 2011

Journal of Biological Chemistry

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“…Regarding the three consensus sites in CXCR-2, it is therefore most probable that Asn 186 , being spaced from the adjacent transmembrane domain by only seven amino acid residues, is not occupied by a carbohydrate moiety. In addition, glycosylation at Asn 186 appears the more unlikely because of the presence of a proline residue in position 189; this kind of amino acid is generally thought to prevent the attachment of sugars when directly preceding or following the glycosylation sequon (33). Hence, Asn 17 and Asn 197 , both located more distal from the membrane, are the most probable candidates to carry the two 9-kDa oligosaccharides in cell membrane-expressed CXCR-2.…”

Section: Discussionmentioning

confidence: 99%

Identification of Distinct Surface-Expressed and Intracellular CXC-Chemokine Receptor 2 Glycoforms in Neutrophils:N-Glycosylation Is Essential for Maintenance of Receptor Surface Expression

Ludwig

Ehlert

Flad

et al. 2000

The Journal of Immunology

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The G protein-coupled CXC-chemokine receptor CXCR-2 mediates activation of neutrophil effector functions in response to multiple ligands, including IL-8 and neutrophil-activating peptide 2 (NAP-2). Although CXCR-2 has been successfully cloned and expressed in several cell lines, the molecular properties of the native neutrophil-expressed receptor have remained largely undefined. Here we report on the identification and characterization of distinct CXCR-2 glycoforms and their subcellular distribution in neutrophils. Immunoprecipitation and Western blot analyses of surface-expressed receptors covalently linked to IL-8 or NAP-2 as well as in their unloaded state revealed the occurrence of a single CXCR-2 variant with an apparent size of 56 kDa. According to deglycosylation experiments surface-expressed CXCR-2 carries two N-linked 9-kDa carbohydrate moieties that are both of complex structure. In addition, two other CXCR-2 variants of 38 and 40 kDa were found to occur exclusively intracellular and to carry N-glycosylations of high mannose or hybrid type. These receptors did not participate in ligand-induced receptor trafficking, while surface-expressed CXCR-2 was internalized and re-expressed following stimulation with NAP-2. By enzymatic removal of one 9-kDa carbohydrate moiety in surface-expressed CXCR-2 we can show that neither NAP-2-induced trafficking nor signaling of the receptor is dependent on its full glycosylation. Instead, glycosylation was found to protect CXCR-2 from proteolytic attack, as even partial deglycosylation is associated with serine protease-mediated disappearance of the receptor from the neutrophil surface. Thus, although not directly involved in signaling, glycosylation appears to be required to maintain neutrophil responsiveness to CXC-chemokines during inflammation.

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“…E-selectin is highly glycosylated and N-linked carbohydrate accounts for a third of its molecular weight [2]. N-linked glycosylation influences many properties of proteins, including folding, transport, activity, stability and antigenicity [9][10][11]. The role of N-glycosylation of E-selectin has previously been studied using HUVEC stimulated with IL-1 and grown in the presence of glycosylation inhibitors.…”

Section: Introductionmentioning

confidence: 99%

Interferon‐γ Enhancement of E‐Selectin Expression on Endothelial Cells is Inhibited by Monensin

1997

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Strindhall J, Lundblad A, Påhlsson P. Interferon-g Enhancement of E-Selectin Expression on Endothelial Cells is Inhibited by Monensin. Scand J Immunol 1997;46:338-343 The expression of E-selectin reaches a maximum 4-6 h after stimulation of human umbilical vein endothelial cells (HUVEC) in vitro with tumour necrosis factor-a (TNF-a) and then declines to basal level within 24 h. If interferon-g (IFN-g) is added to the cell culture medium together with TNF-a the surface expression of Eselectin is augmented and prolonged in a synergistic way. The aim of the present study was to investigate if altered protein glycosylation could explain the IFN-g induced persistent surface expression of E-selectin. SDS-PAGE analysis of HUVEC glycoproteins, metabolically radiolabelled in the carbohydrate portion, indicated that addition of IFN-g produced an altered protein glycosylation. Lectin blot analysis using the Sambucus nigra agglutinin lectin also indicated differences in protein glycosylation when HUVEC were incubated with IFN-g/TNF-a compared to TNF-a alone. The kinetics of surface expression of E-selectin were measured using a cell ELISA assay. When HUVEC were incubated with monensin, a potent inhibitor of late Golgi function, together with both TNF-a and IFN-g, the additive effect of IFN-g on E-selectin expression was almost abolished. Since monensin is known to affect glycosylation processing, this experiment suggested that the IFN-g induced change in protein glycosylation might induce the prolonged surface expression of Eselectin. However, when HUVEC were cultured with IFN-g/TNF-a in the presence of several different inhibitors of N-glycosylation processing, no significant effect of E-selectin expression was observed. Regulation of adhesion molecule expression after activation of endothelial cells is likely to play a pivotal role for the inflammatory response. Further studies are needed to understand the mechanisms underlying this regulation.

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Concepts and principles of glycobiology

Cited by 201 publications

References 5 publications

N-Linked Glycosylation Regulates Human Proteinase-activated Receptor-1 Cell Surface Expression and Disarming via Neutrophil Proteinases and Thermolysin

N-Linked Glycosylation Regulates Human Proteinase-activated Receptor-1 Cell Surface Expression and Disarming via Neutrophil Proteinases and Thermolysin

Identification of Distinct Surface-Expressed and Intracellular CXC-Chemokine Receptor 2 Glycoforms in Neutrophils:N-Glycosylation Is Essential for Maintenance of Receptor Surface Expression

Interferon‐γ Enhancement of E‐Selectin Expression on Endothelial Cells is Inhibited by Monensin

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