Quantitative Glycomics of Human Whole Serum Glycoproteins Based on the Standardized Protocol for Liberating N-Glycans

Kita, Yoko; Miura, Yoshiaki; Furukawa, Yoshihiro; Nakano, Mika; Shinohara, Yasuro; Ohno, Masahiro; Takimoto, Akio; Nishimura, Shin‐Ichiro

doi:10.1074/mcp.t600063-mcp200

Cited by 107 publications

(90 citation statements)

References 44 publications

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“…In brief, IgG samples were reductively alkylated under the presence of detergent and then digested with trypsin and peptide N-glycosidase F (Roche Diagnostics, Penzberg, Germany) (43). The digested samples were mixed with a novel hydrazide-functionalized glycoblotting polymer (42), and sialic acids were methyl-esterified to render sialylated oligosaccharides chemically equivalent to neutral oligosaccharides (44).…”

Section: Analysis Of Oligosaccharide Structuresmentioning

confidence: 99%

Lack of Galactosylation Enhances the Pathogenic Activity of IgG1 but Not IgG2a Anti-Erythrocyte Autoantibodies

Ito

Furukawa

Yamada

et al. 2014

The Journal of Immunology

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IgG bears asparagine-linked oligosaccharide side chains in the Fc region. Variations in their extent of galactosylation and sialylation could modulate IgG Fc-dependent effector functions, and hence Ab activity. However, it has not yet been clarified whether the pathogenic potential of IgG autoantibodies is consistently enhanced by the absence of galactose residues per se or the lack of terminal sialylation, which is dependent on galactosylation. Moreover, it remains to be defined whether the increased pathogenicity of agalactosylated IgG is related to activation of the complement pathway by mannose-binding lectin, as suggested by in vitro studies. Using a murine model of autoimmune hemolytic anemia, we defined the contribution of galactosylation or sialylation to the pathogenic activity of IgG1 and IgG2a anti-erythrocyte class-switch variants of 34-3C monoclonal autoantibody. We generated their degalactosylated or highly sialylated glycovariants and compared their pathogenic effects with those of highly galactosylated or desialylated counterparts. Our results demonstrated that lack of galactosylation, but not sialylation, enhanced the pathogenic activity of 34-3C IgG1, but not IgG2a autoantibodies. Moreover, analysis of in vivo complement activation and of the pathogenic activity in mice deficient in C3 or IgG FcRs excluded the implication of mannose-binding lectin-mediated complement activation in the enhanced pathogenic effect of agalactosylated IgG1 anti-erythrocyte autoantibodies.

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Section: Analysis Of Oligosaccharide Structuresmentioning

confidence: 99%

Lack of Galactosylation Enhances the Pathogenic Activity of IgG1 but Not IgG2a Anti-Erythrocyte Autoantibodies

Ito

Furukawa

Yamada

et al. 2014

The Journal of Immunology

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“…50,51 Briefly, IgG samples were reductively alkylated under the presence of detergent, and then digested with trypsin and peptide N-glycosidase F (Roche Diagnostics, Penzberg, Germany). 52 The digested sample was mixed with a novel hydrazide-functionalized glycoblotting polymer 51 and after washing of the unbound substances, sialic acids were methyl-esterified to render sialylated oligosaccharides chemically equivalent to neutral oligosaccharides. 53 The IgG oligosaccharides were finally recovered as derivatives of aoWR (Na-((aminooxy)acetyl)tryptophanylarginine methyl ester), an oligosaccharide labeling reagent that allows highly sensitive detection on MS. 54 The recovered glycans were subjected to MALDI-TOF MS using an Ultraflex II mass spectrometer (Bruker Daltonik, Bremen, Germany) controlled by the FlexControl 2.0 software package.…”

Section: Analyses Of Oligosaccharide Structuresmentioning

confidence: 99%

Sialylation Determines the Nephritogenicity of IgG3 Cryoglobulins

Otani

Kuroki

Kikuchi

et al. 2012

Journal of the American Society of Nephrology

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Monoclonal 6-19 IgG3 anti-IgG2a rheumatoid factor derived from lupus-prone MRL-Fas lpr mice can induce GN and cryoglobulinemia, but the features that confer nephritogenic potential are not completely understood. Asparagine-linked oligosaccharide chains of 6-19 IgG3 mAb are poorly galactosylated and hardly sialylated, possibly contributing to the pathogenic potential of 6-19 IgG3 rheumatoid factors. Here, we used the 6-19 model of cryoglobulin-associated GN to define the relative contributions of galactosylation and sialylation, in relation to cryoglobulin activity, to the nephritogenic potential of IgG3 antibodies. We generated one highly sialylated and two distinct more galactosylated 6-19 IgG3 rheumatoid factor variants. Although the mere extent of galactosylation had no effect on either the cryogenic and nephritogenic activities of 6-19 IgG3 rheumatoid factor, terminal sialylation attenuated the nephritogenic potential of 6-19 IgG3 by limiting its cryoglobulin activity. These data suggest a protective role of IgG sialylation against the development of cryoglobulin-mediated GN, highlighting the anti-inflammatory activity of sialylated IgG antibodies. Cryoglobulins are a heterogeneous group of Igs that precipitate at temperatures ,37°C, with resolution upon warming. 1,2 Most cryoglobulins are either monoclonal Ig or Ig complexes in which one component, usually IgM, has rheumatoid factor (RF) activity. 1,2 Monoclonal cryoglobulins are mainly associated with various lymphoproliferative disorders, whereas mixed cryoglobulins are often found in sera of patients with autoimmune diseases, such as SLE and rheumatoid arthritis, or with chronic infectious diseases. The presence of cryoglobulins can result in a wide range of vascular, renal, and neurologic complications, likely depending on their concentration, their temperature-dependent solubility behavior, and the nature and type of proteins involved. 2 Antibodies of the IgG3 subclass in humans and mice have the unique physicochemical property that allows them to self-associate via Fc-Fc interactions and to display cryoglobulin activity, independently of their specificities. [3][4][5][6][7][8] Nucleotide sequence analysis of the variable regions of cryogenic and noncryogenic IgG3 mAbs, in combination with the assessment of mutant antibodies, showed that cryoglobulin activity of IgG3 was associated with the presence of more positively charged amino-acid residues at positions 6 and 23 of the heavy-chain variable domain. 9,10 Moreover, structural analysis of asparagine-linked biantennary complex-type oligosaccharide chains (N-glycans) attached to the CH2 domain of different IgG3 mAbs revealed an inverse correlation

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“…In brief, IgG samples were reductively alkylated under the presence of detergent, then digested successively with trypsin and peptide N-glycosidase F (Roche Diagnostics), as previously described (19). The digested sample was mixed with a novel hydrazide-functionalized glycoblotting polymer (18) and, following washing of the unbound substances (e.g., peptides, detergent, enzymes), sialic acids were methyl-esterified to render sialylated oligosaccharides chemically equivalent to neutral oligosaccharides, as described (20).…”

Section: Analysis Of Oligosaccharide Structuresmentioning

confidence: 99%

Impact of a Three Amino Acid Deletion in the CH2 Domain of Murine IgG1 on Fc-Associated Effector Functions

Baudino

Nimmerjahn

Shinohara

et al. 2008

The Journal of Immunology

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Four murine IgG subclasses display markedly different Fc-associated effector functions because of their differential binding to three activating IgG Fc receptors (FcγRI, FcγRIII, and FcγRIV) and C1q. Previous analysis of IgG subclass switch variants of 34-3C anti-RBC monoclonal autoantibodies revealed that the IgG1 subclass, which binds only to FcγRIII and fails to activate complement, displayed the poorest pathogenic potential. This could be related to the presence of a three amino acid deletion at positions 233–235 in the CH2 domain uniquely found in this subclass. To address this question, IgG1 insertion and IgG2b deletion mutants at positions 233–235 of 34-3C anti-RBC Abs were generated, and their ability to initiate effector functions and their pathogenicity were compared with those of the respective wild-type Abs. The insertion of amino acid residues at positions 233–235 enabled the IgG1 subclass to bind FcγRIV but did not improve the binding to C1q. Accordingly, its pathogenicity was enhanced but still inferior to that of IgG2b. In contrast, the IgG2b deletion mutant lost its ability to bind to FcγRIV and activate complement. Consequently, its pathogenicity was markedly diminished to a level comparable to that of IgG1. Our results demonstrated that the initiation of FcγR- and complement-mediated effector functions of IgG2b was profoundly affected by the three amino acid deletion at positions 233–235, but that this natural three amino acid deletion could only partially explain the poor binding of IgG1 to FcγRIV and C1q. This indicates the lack in the IgG1 subclass of as yet unknown motifs promoting efficient interaction with FcγRIV and C1q.

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Quantitative Glycomics of Human Whole Serum Glycoproteins Based on the Standardized Protocol for Liberating N-Glycans

Cited by 107 publications

References 44 publications

Lack of Galactosylation Enhances the Pathogenic Activity of IgG1 but Not IgG2a Anti-Erythrocyte Autoantibodies

Lack of Galactosylation Enhances the Pathogenic Activity of IgG1 but Not IgG2a Anti-Erythrocyte Autoantibodies

Sialylation Determines the Nephritogenicity of IgG3 Cryoglobulins

Impact of a Three Amino Acid Deletion in the CH2 Domain of Murine IgG1 on Fc-Associated Effector Functions

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