Development of a combined chemical and enzymatic approach for the mass spectrometric identification and quantification of aberrant N-glycosylation

Chen, Rui; Wang, Fangjun; Tan, Ye-Xiong; Sun, Zhen; Song, Chunxia; Ye, Mingliang; Wang, Hongyang; Zou, Hanfa

doi:10.1016/j.jprot.2011.12.015

Cited by 35 publications

(29 citation statements)

References 26 publications

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“…Prior work has quantified only the core-fucosylated peptides, but not the non-core-fucosylated counterparts. [22] Hence, it was unclear if changes occurred in the protein amount or extent of core-fucosylation. The work reported in this study quantifies the frequency of core-fucosylation without being influenced by the underlying protein amount.…”

Section: Resultsmentioning

confidence: 99%

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Label‐free relative quantification of alpha‐2‐macroglobulin site‐specific core‐fucosylation in pancreatic cancer by LC‐MS/MS

Lin

Yin

et al. 2013

Electrophoresis

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We describe a label-free relative quantification LC-MS/MS method for core-fucosylationin alpha-2-macroglobulin (A2MG) immunoprecipitated from human sera. The method utilizes endoglycosidase F partial deglycosylation to reduce glycosylation microheterogeneity, while retaining the innermost N-acetylglucosamine (GlcNAc) and core fucose. Precursor ion peak areas of partially deglycosylated peptides were obtained and site-specific core-fucosylation ratios based on the peak areas of core-fucosylated and non-fucosylated counterparts were calculated and evaluated for assay development. This assay was applied in a preliminary study of sera samples from normal controls and patients with pancreatic diseases, including pancreatic cancer and chronic pancreatitis. A2MG fucosylation levels at site N396 and N1424 were found to decrease in both chronic pancreatitis and pancreatic cancer compared to normal controls. The two sites were identified by two peptides and their core-fucosylation ratios were found to be internally consistent. This method provides a platform to quantify fucosylation levels and can be used to study site-specific core-fucosylation aberrations in other glycoproteins for other diseases.

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Section: Resultsmentioning

confidence: 99%

“…Quantitative analysis of serum protein core-fucosylation level changes as a potential hepatocellular carcinoma marker has been performed using precursor intensity-based quantification with differential dimethylation. [22]…”

Section: Introductionmentioning

confidence: 99%

Label‐free relative quantification of alpha‐2‐macroglobulin site‐specific core‐fucosylation in pancreatic cancer by LC‐MS/MS

Lin

Yin

et al. 2013

Electrophoresis

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“…In this way, CID and ETD were not simply alternated during the entire LC-MS/MS run; rather, ETD was only carried out if the CID spectrum yielded evidence that a given precursor ion was a glycopeptide. CID neutral loss products can also be used to trigger ETD for only those analytes likely to be glycosylated [109]. Although not widely applied to glycopeptide analysis at this writing, we note that some previously reported hybrid MS/MS approaches have significant potential to extend the usefulness of MS/MS experiments that yield complementary structural information for glycopeptides [110][111][112][113][114][115].…”

Section: Use Of Complementary Ms/ms Methodsmentioning

confidence: 99%

Engaging Challenges in Glycoproteomics: Recent Advances in MS-Based Glycopeptide Analysis

2015

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The proteomic analysis of glycosylation is uniquely challenging. The numerous and varied biological roles of protein-linked glycans have fueled a tremendous demand for technologies that enable rapid, in-depth structural examination of glycosylated proteins in complex biological systems. In turn, this demand has driven many innovations in wide ranging fields of bioanalytical science. This review will summarize key developments in glycoprotein separation and enrichment, glycoprotein proteolysis strategies, glycopeptide separation and enrichment, the role of mass measurement accuracy in glycopeptide detection, glycopeptide ion dissociation methods for MS/MS, and informatic tools for glycoproteomic analysis. In aggregate, this selection of topics serves to encapsulate the present status of MS-based analytical technologies for engaging the challenges of glycoproteomic analysis.

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“…Stable-isotope dimethyl labelling has also been widely employed in quantitative glycoproteomics [79][80][81]86]. As summarized in table 3, common glycopeptide enrichment methods include lectin affinity chromatography [79], hydrazide capture chemistry [80], and TiO 2 combined with zwitterionic HILIC (ZIC-HILIC) [81,86].…”

Section: (B) Quantitative Analysis Of Protein Post-translational Modimentioning

confidence: 99%

Stable isotope dimethyl labelling for quantitative proteomics and beyond

Hsu

Chen

2016

Phil. Trans. R. Soc. A.

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Stable-isotope reductive dimethylation, a cost-effective, simple, robust, reliable and easy-to- multiplex labelling method, is widely applied to quantitative proteomics using liquid chromatography-mass spectrometry. This review focuses on biological applications of stable-isotope dimethyl labelling for a large-scale comparative analysis of protein expression and post-translational modifications based on its unique properties of the labelling chemistry. Some other applications of the labelling method for sample preparation and mass spectrometry-based protein identification and characterization are also summarized. This article is part of the themed issue ‘Quantitative mass spectrometry’.

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Development of a combined chemical and enzymatic approach for the mass spectrometric identification and quantification of aberrant N-glycosylation

Cited by 35 publications

References 26 publications

Label‐free relative quantification of alpha‐2‐macroglobulin site‐specific core‐fucosylation in pancreatic cancer by LC‐MS/MS

Label‐free relative quantification of alpha‐2‐macroglobulin site‐specific core‐fucosylation in pancreatic cancer by LC‐MS/MS

Engaging Challenges in Glycoproteomics: Recent Advances in MS-Based Glycopeptide Analysis

Stable isotope dimethyl labelling for quantitative proteomics and beyond

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