The Impact of Glycosylation on the Biological Function and Structure of Human Immunoglobulins

Arnold, James N.; Wormald, Mark R.; Sim, Robert B.; Rudd, Pauline M.; Dwek, Raymond A.

doi:10.1146/annurev.immunol.25.022106.141702

Cited by 1,173 publications

(1,129 citation statements)

References 135 publications

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“…This method can be used for relative quantitation of oligosaccharides and is as reliable as chromatographic methods for elucidating glycan profiles (24). The major N-glycans found on IgG molecules are core-fucosylated, partially truncated biantennary species, which may carry a sialic acid species and/or a bisecting N-acetylglucosamine (25,26). Defective glycosylation, especially hypogalactosylation of serum IgG antibodies, has been previously reported in ANCA-positive vasculitides, including GPA (27), but to date, the association with clinical activity has been unknown.…”

Section: Discussionmentioning

confidence: 99%

“…IgG antibodies are heterogeneous with respect to the composition of the N297-attached sugar moiety. While the heptameric core, consisting of N-acetylglucosamine and mannose, is constant, a high level of heterogeneity is introduced through various additions of terminal sugar residues, such as sialic acid and galactose, and branching residues, including N-acetylglucosamine and fucose (25). More than 30 different IgG glycosylation variants exist, which remain at a remarkably constant level in the serum of healthy individuals.…”

Section: Discussionmentioning

confidence: 99%

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Sialylation levels of anti-proteinase 3 antibodies are associated with the activity of granulomatosis with polyangiitis (Wegener's)

Espy

Morelle²,

Kavian

et al. 2011

Arthritis & Rheumatism

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Objective. To investigate whether the glycosylation and sialylation levels of anti-proteinase 3 (anti-PR3) antibodies could affect their pathogenicity, and whether these levels could be correlated with the activity of granulomatosis with polyangiitis (Wegener's) (GPA).Methods. Forty-two serum samples positive for anti-PR3 antibodies from 42 patients with active or weakly active/inactive GPA were included. Anti-PR3 antibodies were assayed by enzyme-linked immunosorbent assay, and their levels of glycosylation and sialylation were assessed by enzyme-linked lectin assay. The glycosylation and sialylation levels of IgG purified from the serum of healthy donors and patients with active, remitted, or weakly active disease were assessed by permethylation and mass spectrometry analysis of glycans, following neuraminidase digestion. The neutrophil oxidative burst induced by purified IgG was assayed by spectrofluorimetry.Results. The mean sialylation ratio of anti-PR3 antibodies was significantly lower in patients with active disease than in patients with weakly active or inactive disease, and this was inversely correlated with the Birmingham Vasculitis Activity Score (BVAS) (P < 0.0001). Similar results were obtained using the BVAS/ GPA. The area under the receiver operating characteristic curve for the sialylation ratio of anti-PR3 antibodies, as a test to determine the activity of GPA, was 0.82 (P ‫؍‬ 0.0006). The characterization of N-glycans showed a decrease in 2,6-linked sialylated N-glycans and an increase in dHex 1 Hex 3 HexNAc 4 (mass/charge 1,836) agalactosylated structures in purified IgG from patients with active disease compared with controls. The anti-PR3 antibody-induced oxidative burst of neutrophils was inversely correlated with the sialylation levels of anti-PR3 IgG.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Sialylation levels of anti-proteinase 3 antibodies are associated with the activity of granulomatosis with polyangiitis (Wegener's)

Espy

Morelle²,

Kavian

et al. 2011

Arthritis & Rheumatism

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“…The impact of glycosylation, one of the most important post-translational modifications, on the structure and biological properties of glycoproteins has been well documented [1,2]. IgG molecules are mainly glycosylated at Asn-297 of the CH2 domain within the Fc region [3,4], with variable galactosylation but limited sialylation.…”

Section: Introductionmentioning

confidence: 99%

Endoglycosidase treatment abrogates IgG arthritogenicity: Importance of IgG glycosylation in arthritis

et al. 2007

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The glycosylation status of IgG has been implicated in the pathology of rheumatoid arthritis. Earlier, we reported the identification of a novel secreted endo-b-Nacetylglucosaminidase (EndoS), secreted by Streptococcus pyogenes that specifically hydrolyzes the b-1,4-di-N-acetylchitobiose core of the asparagine-linked glycan of human IgG. Here, we analyzed the arthritogenicity of EndoS-treated collagen type II (CII)-specific mouse mAb in vivo. Endoglycosidase treatment of the antibodies inhibited the induction of arthritis in (BALB/c Â B10.Q) F1 mice and induced a milder arthritis in B10.RIII mice as compared with the severe arthritis induced by non-treated antibodies. Furthermore, EndoS treatment did not affect the binding of IgG to CII and their ability to activate complement, but it resulted in reduced IgG binding to FccR and disturbed the formation of stable immune complexes. Hence, the asparagine-linked glycan on IgG plays a crucial role in the development of arthritis. IntroductionThe impact of glycosylation, one of the most important post-translational modifications, on the structure and biological properties of glycoproteins has been well documented [1,2]. IgG molecules are mainly glycosylated at Asn-297 of the CH2 domain within the Fc region [3,4], with variable galactosylation but limited sialylation. The remaining glycosylation occurs in the hypervariable regions of the Fab region, with the position and frequency of occurrence being dependent on the presence of the consensus sequence Asn-Xaa-Thr/ Ser for N-glycosylation, and is characterized by a high incidence of sialylated structures. Murine IgG contain 2.3 asparagine-linked (N-linked) biantennary oligosaccharide chains per molecule [5], and human IgG contain 2.8 [6]. The minimal oligosaccharide structure is a hexasaccharide (GlcNAc2Man3GlcNAc) with variable sugar residues attached, which results in the generation of the multiple glycoforms. About 30 variants of biantennary chains occur, resulting in Clinical immunology

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“…Protein glycosylation is a biologically significant and complex post-translational modification, involved in cell-cell and receptor-ligand interactions (1)(2)(3)(4). In fact, clinical biomarkers and therapeutic targets are often glycoproteins (5)(6)(7)(8)(9).…”

mentioning

confidence: 99%

Identification and Quantification of Glycoproteins Using Ion-Pairing Normal-phase Liquid Chromatography and Mass Spectrometry

Ding

Nothaft

Szymanski

et al. 2009

Molecular & Cellular Proteomics

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Glycoprotein structure determination and quantification by MS requires efficient isolation of glycopeptides from a proteolytic digest of complex protein mixtures. Here we describe that the use of acids as ion-pairing reagents in normal-phase chromatography (IP-NPLC) considerably increases the hydrophobicity differences between nonglycopeptides and glycopeptides, thereby resulting in the reproducible isolation of N-linked high mannose type and sialylated glycopeptides from the tryptic digest of a ribonuclease B and fetuin mixture. The elution order of nonglycopeptides relative to glycopeptides in IP-NPLC is predictable by their hydrophobicity values calculated using the Wimley-White water/octanol hydrophobicity scale. Olinked glycopeptides can be efficiently isolated from fetuin tryptic digests using IP-NPLC when N-glycans are first removed with PNGase. IP-NPLC recovers close to 100% of bacterial N-linked glycopeptides modified with non-sialylated heptasaccharides from tryptic digests of periplasmic protein extracts from Campylobacter jejuni 11168 and its pglD mutant. Label-free nano-flow reversed-phase LC-MS is used for quantification of differentially expressed glycopeptides from the C. jejuni wildtype and pglD mutant followed by identification of these glycoproteins using multiple stage tandem MS. This method further confirms the acetyltransferase activity of PglD and demonstrates for the first time that heptasaccharides containing monoacetylated bacillosamine are transferred to proteins in both the wild-type and mutant strains. We believe that IP-NPLC will be a useful tool for quantitative glycoproteomics. Molecular & Cellular Proteomics 8:2170 -2185, 2009.Protein glycosylation is a biologically significant and complex post-translational modification, involved in cell-cell and receptor-ligand interactions (1-4). In fact, clinical biomarkers and therapeutic targets are often glycoproteins (5-9). Comprehensive glycoprotein characterization, involving glycosylation site identification, glycan structure determination, site occupancy, and glycan isoform distribution, is a technical challenge particularly for quantitative profiling of complex protein mixtures (10 -13). Both N-and O-glycans are structurally heterogeneous (i.e. a single site may have different glycans attached or be only partially occupied). Therefore, the MS 1 signals from glycopeptides originating from a glycoprotein are often weaker than from non-glycopeptides. In addition, the ionization efficiency of glycopeptides is low compared with that of non-glycopeptides and is often suppressed in the presence of non-glycopeptides (11-13). When the MS signals of glycopeptides are relatively high in simple protein digests then diagnostic sugar oxonium ion fragments produced by, for example, front-end collisional activation can be used to detect them. However, when peptides and glycopeptides coelute, parent ion scanning is required to selectively detect the glycopeptides (14). This can be problematic in terms of sensitivity, especially for detecting glycopep...

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The Impact of Glycosylation on the Biological Function and Structure of Human Immunoglobulins

Cited by 1,173 publications

References 135 publications

Sialylation levels of anti-proteinase 3 antibodies are associated with the activity of granulomatosis with polyangiitis (Wegener's)

Sialylation levels of anti-proteinase 3 antibodies are associated with the activity of granulomatosis with polyangiitis (Wegener's)

Endoglycosidase treatment abrogates IgG arthritogenicity: Importance of IgG glycosylation in arthritis

Identification and Quantification of Glycoproteins Using Ion-Pairing Normal-phase Liquid Chromatography and Mass Spectrometry

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