The Glycosylation of the Complement Regulatory Protein, Human Erythrocyte CD59

Rudd, Pauline M.; Morgan, B. Paul; Wormald, Mark R.; Harvey, David J.; Berg, Carmen W. van den; Davis, Simon J.; Ferguson, Michael A. J.; Dwek, Raymond A.

doi:10.1074/jbc.272.11.7229

Cited by 164 publications

(110 citation statements)

References 55 publications

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“…39−41 The number of glycans observed in IgG contrasts with the large heterogeneity found in other glycoproteins of the immune system. 42 For instance, a single GPI-anchored glycoprotein, CD59, has been reported to contain over 120 glycoforms. 42 CE-MS Analysis of N-Glycans from rHuEPO.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

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Simple Capillary Electrophoresis–Mass Spectrometry Method for Complex Glycan Analysis Using a Flow-Through Microvial Interface

et al. 2014

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A flow-through microvial is used to interface capillary electrophoresis and mass spectrometry (CE-MS) to develop a method for simultaneous profiling both neutral and sialylated glycans without derivatization or labeling. The CE separation was performed at near-zero electroosmotic flow in a capillary with neutral, hydrophilic coating, using 50 mM ammonium acetate in 20% methanol (pH 3.1) as the background electrolyte. The method was optimized with reversed CE polarity and negative ion ESI-MS. Enzymatically released N-glycans from human immunoglobulin G (IgG) were used as the test sample. The approach was also used to study the more complex N-glycans from recombinant human erythropoietin (rHuEPO) expressed in Chinese hamster ovary (CHO) cells. Glycoscreening of rHuEPO was performed using a triple quadrupole MS and an ultrahigh resolution TOF-MS. The high sensitivity and high mass accuracy of the TOF-MS revealed the presence of more than 70 glycans. Three mono-and di-sialylated tetra-antennary N-glycans and one mono-sialylated tri-antennary N-glycan of rHuEPO are reported for the first time. Further glycan heterogeneity was identified of the highly sialylated N-glycans of rHuEPO by extensive acetylation, Neu5Ac/Neu5Gc variation and the presence of N-acetyl-lactosamine repeats. For comparative purposes, porous graphitic carbon-based LC-MS/MS was also used to glycoprofile rHuEPO. This work demonstrates the potential of CE-MS to provide a comprehensive glycosylation profile with detailed features of the secondary glycan modifications. The CE-MS based method eliminates the need to label the N-glycans, as well as the requirement to desialylate before analysis, and could complement other established techniques for glycan characterization of therapeutic glycoproteins.

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Section: ■ Results and Discussionmentioning

confidence: 99%

“…42 For instance, a single GPI-anchored glycoprotein, CD59, has been reported to contain over 120 glycoforms. 42 CE-MS Analysis of N-Glycans from rHuEPO. Previous CE-MS studies on N-glycosylation of rHuEPO were performed using bare-fused silica capillaries in basic background electrolyte (BGE) conditions.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Simple Capillary Electrophoresis–Mass Spectrometry Method for Complex Glycan Analysis Using a Flow-Through Microvial Interface

et al. 2014

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“…An aliquot of the aqueous sample (ϳ25 pmol) was mixed with acetonitrile in the ratio 30:70 and applied to the NP-HPLC column. Conditions for the digestions are as described previously (42). The cleavage positions for the exoglycosidases are shown in Fig.…”

Section: Methodsmentioning

confidence: 99%

“…6 shows the digestion of the total glycan pool from IgA1 with different exoglycosidase arrays monitored by NP-HPLC. The assignment of each of the main peaks in the IgA1 glycan pool was confirmed by following its predicted elution position through each of the enzyme digestions (Table II) as described (42). Predicted elution positions were based upon predetermined incremental values for the monosaccharide additions to standard glycan cores (40) and the known specificity of the enzymes (Fig.…”

Section: Iga1 Glycosylation and N-glycan Function In Fc␣r Bindingmentioning

confidence: 99%

The Glycosylation and Structure of Human Serum IgA1, Fab, and Fc Regions and the Role of N-Glycosylation on Fcα Receptor Interactions

Mattu¹,

Pleass²,

Willis³

et al. 1998

Journal of Biological Chemistry

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378

363

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“…The glycosylation of the complcn3cnt rcgulatory protein human crythrocxtc (:1)59 has been anah'zcd [36]. 'l'his ccll-surfacc glycoprotcin contains .V-glycans, O-glycans and a ~lycosylphosphatidylinositol anchor.…”

Section: New Protein Glycosylation Sitesmentioning

confidence: 99%

Novel forms of protein glycosylation

Vliegenthart

Casset

1998

Current Opinion in Structural Biology

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A large number of new glycans, derived from glycoproteins, has been characterized in the past few years. O-linked fucose was found in epidermal growth factor-like domains of several proteins. For the N-linked glycans of Helix pomatia hemocyanin, novel types of antennae were identified. The positions of noncarbohydrate substituents were established in N-glycans. C-mannosylation of a tryptophan residue was discovered in human ribonuclease 2 and is the first example of C-glycosylation in glycoproteins. Addresses

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The Glycosylation of the Complement Regulatory Protein, Human Erythrocyte CD59

Cited by 164 publications

References 55 publications

Simple Capillary Electrophoresis–Mass Spectrometry Method for Complex Glycan Analysis Using a Flow-Through Microvial Interface

Simple Capillary Electrophoresis–Mass Spectrometry Method for Complex Glycan Analysis Using a Flow-Through Microvial Interface

The Glycosylation and Structure of Human Serum IgA1, Fab, and Fc Regions and the Role of N-Glycosylation on Fcα Receptor Interactions

Novel forms of protein glycosylation

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