Dephosphorylation of intact glycoprotein to greatly improve digestion efficiency coupled with matrix-assisted laser desorption/ionization–Fourier transform ion cyclotron resonance mass spectrometric analysis

Li, Fenjie; Wang, Xinli; Liu, Yujie; Liu, Hui; Li, Zhili

doi:10.1016/j.aca.2013.05.044

Cited by 6 publications

(9 citation statements)

References 35 publications

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“…An approach for the simultaneous enrichment and identification of phosphopeptides and sialylated glycopeptides has been developed . It is worth mentioning that the enzymatic digestion efficiency of phosphorylated glycoproteins can be improved by removing phosphoric acid groups . In addition, it has been shown that the combination of fractionation and TiO 2 chromatography exhibits better enrichment capacity for sialylated glycopeptides compared with only lectin or TiO 2 chromatography …”

Section: Glycopeptidesmentioning

confidence: 99%

“…112 It is worth mentioning that the enzymatic digestion efficiency of phosphorylated glycoproteins can be improved by removing phosphoric acid groups. 114 In addition, it has been shown that the combination of fractionation and TiO 2 chromatography exhibits better enrichment capacity for sialylated glycopeptides compared with only lectin or TiO 2 chromatography. 110 A method called reverse glycoblotting, which allows quantitative and specific enrichment of various compounds bearing one or more sialic acid residues, has been developed for the selective isolation of sialylated glycopeptides.…”

Section: Ce-ms For Glycoprotein Analysismentioning

confidence: 99%

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Mass spectrometry for protein sialoglycosylation

Zhang

Wang

et al. 2017

Mass Spectrometry Reviews

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Sialic acids are a family of structurally unique and negatively charged nine-carbon sugars, normally found at the terminal positions of glycan chains on glycoproteins and glycolipids. The glycosylation of proteins is a universal post-translational modification in eukaryotic species and regulates essential biological functions, in which the most common sialic acid is N-acetyl-neuraminic acid (2-keto-5-acetamido-3,5-dideoxy-D-glycero-D-galactononulopyranos-1-onic acid) (Neu5NAc). Because of the properties of sialic acids under general mass spectrometry (MS) conditions, such as instability, ionization discrimination, and mixed adducts, the use of MS in the analysis of protein sialoglycosylation is still challenging. The present review is focused on the application of MS related methodologies to the study of both N- and O-linked sialoglycans. We reviewed MS-based strategies for characterizing sialylation by analyzing intact glycoproteins, proteolytic digested glycopeptides, and released glycans. The review concludes with future perspectives in the field.

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Section: Glycopeptidesmentioning

confidence: 99%

Section: Ce-ms For Glycoprotein Analysismentioning

confidence: 99%

Mass spectrometry for protein sialoglycosylation

Zhang

Wang

et al. 2017

Mass Spectrometry Reviews

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“…Among these factors are the quantification method (of the purified fraction) and the dye applied for visualization of the spots in gel electrophoresis. Furthermore, glycosylated protein digestion by trypsin is often hampered due to incomplete digestion .…”

Section: Introductionmentioning

confidence: 99%

Quantification, 2DE analysis and identification of enriched glycosylated proteins from mouse muscles: Difficulties and alternatives

et al. 2015

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One of the problems with 2DE is that proteins present in low amounts in a sample are usually not detected, since their signals are masked by the predominant proteins. The elimination of these abundant proteins is not a guaranteed solution to achieve the desired results. The main objective of this study was the comparison of common and simple methodologies employed for 2DE analysis followed by MS identification, focusing on a pre-purified sample using a wheat germ agglutinin (WGA) column. Adult male C57Black/Crj6 (C57BL/6) mice were chosen as the model animal in this study; the gastrocnemius muscles were collected and processed for the experiments. The initial fractionation with succinylated WGA was successful for the elimination of the most abundant proteins. Two quantification methods were employed for the purified samples, and bicinchoninic acid (BCA) was proven to be most reliable for the quantification of glycoproteins. The gel staining method, however, was found to be decisive for the detection of specific proteins, since their structures affect the interaction of the dye with the peptide backbone. The Coomassie Blue R-250 dye very weakly stained the gel with the WGA purified sample. When the same gel was stained with silver nitrate, however, MS could positively assign 12 new spots. The structure of the referred proteins was not found to be prone to interaction with Coomassie blue.

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“…Increased expression of SA on the surface of cancer cells may lead to its definition as potential therapeutic targets . However, because of a nontemplate driven biosynthesis of the numerous and diverse glycan structures and high heterogeneity of sialylation sites, as well as their low abundance, low ionization efficiency, and losses of SA residues and water molecule during analytical processes, , these factors have made glycomic analysis to face great challenge …”

mentioning

confidence: 99%

“…A number of efforts to enrich and detect sialoglycopeptides and/or glycopeptides from complex biological samples have been made, such as lectin, − serotonin-bonded silica, hydrazine chemistry, , titanium dioxide chromatography, ,, hydrophilic interaction chromatography, and strong cation exchange chromatography . However, some shortages of these strategies, including low specificity, time-consuming, and potential losses of SA residues and water molecule from sialoglycopeptides during analytical processes, have limited their applications in deciphering correctly sialylation of glycoproteins.…”

mentioning

confidence: 99%

Kapok Fiber: A Natural Biomaterial for Highly Specific and Efficient Enrichment of Sialoglycopeptides

Liu

Zhang

et al. 2015

Anal. Chem.

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Cancer development and chronic diseases are associated with the overexpression of sialoglycans terminated to the surface proteins and lipids of cancer cells compared with normal cells. The isolation and detection of sialoglycopeptides from complex peptides mixture still remain challenges due to their low abundance, low ionization, and losses of sialic acid residues and water molecule during analytical processes. In this study, kapok fiber, a natural fiber derived from the kapok tree (Bombax ceiba L.), has shown excellent capability to specifically and efficiently enrich sialoglycopeptides, without losses of sialic acid residues and water molecule from sialoglycans. The main components on the surface of kapok fiber are syringyl and guaiacyl lignins which play an important role in isolating sialoglycopeptides from complex peptide mixtures.

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Dephosphorylation of intact glycoprotein to greatly improve digestion efficiency coupled with matrix-assisted laser desorption/ionization–Fourier transform ion cyclotron resonance mass spectrometric analysis

Cited by 6 publications

References 35 publications

Mass spectrometry for protein sialoglycosylation

Mass spectrometry for protein sialoglycosylation

Quantification, 2DE analysis and identification of enriched glycosylated proteins from mouse muscles: Difficulties and alternatives

Kapok Fiber: A Natural Biomaterial for Highly Specific and Efficient Enrichment of Sialoglycopeptides

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