The effect of aglycosylation on the binding of mouse IgG to staphylococcal protein A

Leatherbarrow, Robin J.; Dwek, Raymond A.

doi:10.1016/0014-5793(83)80290-7

Cited by 38 publications

(14 citation statements)

References 30 publications

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“…Treated IgG was applied to a membrane in serial dilutions and probed with radiolabeled, purified protein H. This revealed that protein H bound IgG to the same extent irrespective of EndoS treatment and indicated that the Fc‐binding capacity of protein H is not influenced by EndoS (Figure 8A). This was also the case for staphylococcal protein A, which bound to both intact and EndoS‐treated IgG (data not shown), as previously described for aglycosyl mouse IgG (Leatherbarrow and Dwek, 1983; Nose and Wigzell, 1983).…”

Section: Resultsmentioning

confidence: 99%

EndoS, a novel secreted protein from Streptococcus pyogenes with endoglycosidase activity on human IgG

Collin¹

2001

The EMBO Journal

334

300

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Streptococcus pyogenes is an important human pathogen that selectively interacts with proteins involved in the humoral defense system, such as immunoglobulins and complement factors. In this report we show that S.pyogenes has the ability to hydrolyze the chitobiose core of the asparagine-linked glycan on immunoglobulin G (IgG) when bacteria are grown in the presence of human plasma. This activity is associated with the secretion of a novel 108 kDa protein denoted EndoS. EndoS has endoglycosidase activity on puri®ed soluble IgG as well as IgG bound to the bacterial surface. EndoS is required for the activity on IgG, as an isogenic EndoS mutant could not hydrolyze the glycan on IgG. In addition, we show that the secreted streptococcal cysteine proteinase SpeB cleaves IgG in the hinge region in a papain-like manner. This is the ®rst example of an endoglycosidase produced by a bacterial pathogen that selectively hydrolyzes human IgG, and reveals a novel mechanism which may contribute to S.pyogenes pathogenesis. Keywords: cysteine proteinase/endo-b-N-acetylglucosaminidase/endoglycosidase/IgG/Streptococcus pyogenes IntroductionAn enzyme expressed by Streptococcus pyogenes capable of releasing the terminal sialic acid residues from glycoproteins such as immunoglobulins was ®rst described by Hayano and Tanaka (1967). Other studies have shown that this release of sialic acid from immunoglobulin G (IgG) is due to a neuraminidase activity produced by S.pyogenes, resulting in autoantigenic and nephritogenic properties in patients with glomerulonephritis (Mosquera et al., 1985). Related endoglycosidases found in Staphylococcus aureus have been shown to be cluster-dispersing enzymes involved in cell separation (Sugai et al., 1995), and glycosidases in Streptococcus oralis that sequentially deglycosylate N-linked glycoproteins promote growth of the bacterium (Byers et al., 1999). It has also been suggested that a b-N-acetylglucosaminidase from Streptococcus pneumoniae contributes to pathogenicity, since N-linked sugar residues are common features of several cell-surface proteins in host tissue (Clarke et al., 1995).Oligosaccharide-cleaving enzymes from other species have been studied in detail, and some are standard tools in glycobiology research. For example, endoglycosidase H from Streptomyces plicatus (Trumbly et al., 1985), and endoglycosidases F 1 , F 2 and F 3 from Flavobacterium meningosepticum (Plummer and Tarentino, 1991;Trimble and Tarentino, 1991) are commonly used. These enzymes all belong to family 18 of chitinases (Henrissat, 1991) and catalyze the hydrolysis of the b-1,4-N-acetyl-D-glucosamine linkages in the chitin core of N-linked oligosaccharides in glycoproteins. These enzymes are referred to as true endoglycosidases (Alexander and Elder, 1989;Tarentino and Plummer, 1994). However, a hydrolase isolated from F.meningosepticum, peptide-N-(N-acetylb-glucosaminyl)asparagine amidase (PNGase F), has activity on the pentasaccharide core region of asparagine-linked glycans, and is therefore referred to as a deglycos...

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Section: Resultsmentioning

confidence: 99%

EndoS, a novel secreted protein from Streptococcus pyogenes with endoglycosidase activity on human IgG

Collin¹

2001

The EMBO Journal

334

300

View full text Add to dashboard Cite

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“…Thermal stability and functionality of the CH2 domains of IgG are progressively reduced with successive removal of outer-arm sugar residues [60]. Aglycosylated IgG fails to activate complement [19], is more liable to proteolytic attack [18] and is not recognized by cells expressing FccRI and II receptors [61]. Furthermore, removal of the complete carbohydrate moiety abolished the ability to activate complement and ADCC of a human IgG1 mAb, Campath-1H, but left the antigen and protein A binding activities intact, whereas removal of terminal sialic acid residues through glycopeptidase-F digestion did not affect any of Figure 6.…”

Section: Discussionmentioning

confidence: 99%

“…Modifications in these oligosaccharides affect susceptibility to proteolytic degradation, clearance rate, Ab-dependent cellular cytotoxicity (ADCC), as well as complement activation apart from binding to FcR, monocytes, protein G and C1q/C1 [12][13][14][15][16][17]. However, Fc glycosylation is not required for either protein A binding [18] or recognition of antigen [19][20][21][22]. Recently, de-fucosylation on the N297-linked glycan in the Fc part of the Ab has been shown to increase ADCC activity, indicating the importance of glyco-engineering of Ab for improved clinical efficacy [23].…”

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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“…Aglycosylated IgG, obtained by culturing IgG-producing cell lines in the presence of tunicamycin, has been shown to lack several important biological functions, e.g. binding to monocyte FcyRI receptor (Leatherbarrow et al, 1985;Walker et al, 1989), to display decreased complement activation (Nose & Wigzell, 1983;Leatherbarrow & Dwek, 1983) and to display increased susceptibility to proteolysis (Leatherbarrow & Dwek, 1983). However, glycopeptides isolated after proteolytic digestion of IgG have not been demonstrated to have biological activity, e.g.…”

Section: Introductionmentioning

confidence: 99%

A comparative study of the N-linked oligosaccharide structures of human IgG subclass proteins

Jefferis

Lund

Mizutani

et al. 1990

Biochemical Journal

156

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Quantitative oligosaccharide profiles were determined for each of 18 human IgG paraproteins representing the four subclasses. Each paraprotein exhibits a unique profile that may be substantially different from that observed for polyclonal IgG. The IgG2 and some IgG3 proteins analysed exhibit a predominance of oligosaccharide moieties having galactose on the Man(alpha 1----3) arm rather than the Man(alpha 1----6) arm; it was previously held that galactosylation of the Man(alpha 1----6) arm is preferred, as observed for IgG1, IgG4 and polyclonal IgG. An IgG4 protein is reported that has galactosylated Man(alpha 1----3) and Man(alpha 1----6) arms on both Fc-localized carbohydrate moieties; previous findings suggested that such fully glycosylated structures could not be accommodated within the internal space of the C gamma 2 domains. Unusual monoantennary oligosaccharides present in IgG2 and IgG3 proteins were isolated and their structures determined.

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The effect of aglycosylation on the binding of mouse IgG to staphylococcal protein A

Cited by 38 publications

References 30 publications

EndoS, a novel secreted protein from Streptococcus pyogenes with endoglycosidase activity on human IgG

EndoS, a novel secreted protein from Streptococcus pyogenes with endoglycosidase activity on human IgG

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

A comparative study of the N-linked oligosaccharide structures of human IgG subclass proteins

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