Site-specific N-glycosylation analysis of human plasma ceruloplasmin using liquid chromatography with electrospray ionization tandem mass spectrometry

Harazono, Akira; Kawasaki, Nana; Itoh, Seiga; Hashii, Noritaka; Ishii‐Watabe, Akiko; Kawanishi, Toru; Hayakawa, Tetsuo

doi:10.1016/j.ab.2005.10.036

Cited by 58 publications

(59 citation statements)

References 28 publications

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“…The MS/MS spectrum in Figure 5A displays an abundant ion corresponding to the protonated peptide 173-178, which was formed by the complete loss of the oligosaccharide chain. Remarkably, the MS/MS spectrum of the shorter peptide 173-178 ( Figure 5A The observation of peptide fragments which partially retain N-acetyl glucosamine is in agreement with data published by other groups [15,17], which showed that the GlcNAc was retained on the y-ions. Their data suggests that the formation of such fragments, however, is dependent on the amino acid composition and location of the glycosylation site within the peptide.…”

Section: Peptide Backbone Fragmentation (In the Cid Of Glycoconjugates)supporting

confidence: 78%

“…Under these conditions, however, fragmentation of the glycan was complete and fragment ions due to successive loss of glycans were no longer observed [17]. Other groups used collision energies up to 80 V in order to observe fragmentation of the peptide [15]. The glycosylation site N174 (N556), observed in the chymotryptic peptide 173-178 and in the peptide 171-180, formed by non-specific activity of the enzyme, is occupied with a heterogeneous population of high mannose glycans, ranging from the minimal core structure Man3 up to nine mannoses attached to the chitobiose core (Man9).…”

Section: Peptide Backbone Fragmentation (In the Cid Of Glycoconjugates)mentioning

confidence: 99%

“…Several approaches have become commonly used to address this issue: (i) analysis of the N-linked carbohydrates released from the protein by enzymatic procedures followed by derivatization of the glycan pool [14], or (ii) analysis of the glycopeptides formed by proteolytic degradation of the protein [15,16]. While the former provides a global picture of the glycans decorating the protein, the second strategy discloses information about the carbohydrate attachment site, which can be obtained using various tandem mass spectrometric methods (reviewed in [17]).…”

Section: Introductionmentioning

confidence: 99%

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Site Specific Identification of N-Linked Glycosylation in Proteins by Liquid Chromatography–Electrospray Ionization Tandem Mass Spectrometry

Perdivara

Iacob

Przybylski

et al. 2008

NATO Science for Peace and Security Series A: Chemistry and Biology

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Abstract.Recently, we reported the characterization of the glycans attached at the 11 N-glycosylation sites of Hepatitis C virus E2 envelope glycoprotein by tandem mass spectrometry. Infections caused by Hepatitis C virus represent the main cause of liver diseases such as hepatitis, cirrhosis and hepatocellular carcinoma. The N-linked sugars consist primarily of high mannose glycans, with structures ranging from the minimal core structure, Man3GlcNAc2 (Man3) up to 12 hexose residues attached to the GlcNAc-ß(1-4)-GlcNAc core (depicted as Hex3Man9GlcNAc2). Furthermore, the site N41 (N423) was observed to contain complex type glycans with the structures Man3-GlcNAc and Man3-GlcNAcFuc, in addition to the high mannose population Man3 through Man6, while the site N48 (N430) was occupied exclusively with complex type glycans (Man3-Fuc, Man3-GlcNAcFuc and Man3-GlcNAc2Fuc). The present contribution summarizes our experimental observations upon the factors which may have an impact on the CID tandem mass spectra of glycopeptides.

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Section: Peptide Backbone Fragmentation (In the Cid Of Glycoconjugates)supporting

confidence: 78%

Section: Peptide Backbone Fragmentation (In the Cid Of Glycoconjugates)mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Site Specific Identification of N-Linked Glycosylation in Proteins by Liquid Chromatography–Electrospray Ionization Tandem Mass Spectrometry

Perdivara

Iacob

Przybylski

et al. 2008

NATO Science for Peace and Security Series A: Chemistry and Biology

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“…Several methods are applied to characterize EPO by using high pH anion exchange chromatography (HPAEC-PAD) and NP-HPLC with mass spectrometry. 5 These include the structural characterization of glycoproteins, modifications of glycoproteins, composition analysis of released neutral glycans, and structural analysis of glycans. 6 The higher complexities of peptides and proteins compared to organic low molecular weight drugs and the different ways that can be used to produce biotechnological products necessitates special requirements concerning quality assurance and analytical testing.…”

Section: Introductionmentioning

confidence: 99%

Analysis of the Structure and Stability of Erythropoietin by pH and Temperature Changes using Various LC/MS

Chang¹,

Kim²,

Kim³

2013

Bulletin of the Korean Chemical Society

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The purpose of stability testing is to provide evidence about how the quality of a drug varies with time under the influence of a variety of environmental factors. In this study, erythropoietin (EPO) was analyzed under different pH (pH 3 and pH 9) and temperature (25 o C and 40 o C) conditions according to current Good Manufacturing Practice (cGMP) and International Conference on Harmonisation (ICH) guidelines. The molecular weight difference between intact EPO and deglycosylated EPO was determined by SDS-PAGE, and aggregated forms of EPO under thermal stress and high-pH conditions were investigated by size exclusion chromatography. High pH and high temperature induced increases in dimer and high molecular weight aggregate forms of EPO. UPLC-ESI-TOF-MS was applied to analyze the changed modification sites on EPO. Further, normal-phase high-performance liquid chromatography was performed to identify proposed glycan structures and high pH anion exchange chromatography was carried out to investigate any change in carbohydrate composition. The results demonstrated that there were no changes in modification sites or the glycan structure under severe conditions; however, the number of dimers and aggregates increased at 40 o C and pH 9, respectively.

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“…[5][6][7] In bottom-up MS-based glycosylation analysis, glycoproteins are first digested into short glycopeptides by a protease, and then the resulting glycopeptides are enriched and analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). 8,9 Fragmentation methods in MS/MS determine the patterns of fragment ions observed in MS/MS spectra of glycopeptides.…”

Section: Introductionmentioning

confidence: 99%

Identification of Glycopeptides with Multiple Hydroxylysine O-Glycosylation Sites by Tandem Mass Spectrometry

Zhang

Song

et al. 2015

J. Proteome Res.

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Glycosylation is one of the most common post-translational modifications in proteins, existing in about 50% of mammalian proteins. Several research groups have demonstrated that mass spectrometry is an efficient technique for glycopeptide identi- we present GlycoMID, a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra. GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptidespectrum-matches than other existing tools, including many glycopeptides with two glycosylation sites.

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Site-specific N-glycosylation analysis of human plasma ceruloplasmin using liquid chromatography with electrospray ionization tandem mass spectrometry

Cited by 58 publications

References 28 publications

Site Specific Identification of N-Linked Glycosylation in Proteins by Liquid Chromatography–Electrospray Ionization Tandem Mass Spectrometry

Site Specific Identification of N-Linked Glycosylation in Proteins by Liquid Chromatography–Electrospray Ionization Tandem Mass Spectrometry

Analysis of the Structure and Stability of Erythropoietin by pH and Temperature Changes using Various LC/MS

Identification of Glycopeptides with Multiple Hydroxylysine O-Glycosylation Sites by Tandem Mass Spectrometry

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