Improved online LC-MS/MS identification of O-glycosites by EThcD fragmentation, chemoenzymatic reaction, and SPE enrichment

Yang, Shuang; Wang, Yan; Mann, Matthew; Wang, Qiong; Tian, E; Zhang, Liping; Cipollo, John F.; Hagen, Karl S.; Tabak, Lawrence A.

doi:10.1007/s10719-020-09952-w

Cited by 15 publications

(15 citation statements)

References 37 publications

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“…As shown in Figure 4, proteases can cleave peptides at a variety of amino acid residues, predominantly at the C‐terminus of amino acids. When glycopeptides contain K, R, and histidine (H), they have more positive charges, but when glycopeptides are composed of negatively charged Asp (D) or Glu (E), the total positive charges will decrease when the positive‐mode is applied [47]. Since proteases cleave the glycopeptides at the P1 and P1′ positions, the use of different proteases will result in different glycopeptide compositions (Figure 4A).…”

Section: General Workflow For Analyzing Glycans and Glycopeptidesmentioning

confidence: 99%

Advances in glycopeptide enrichment methods for the analysis of protein glycosylation over the past decade

Zhang

et al. 2022

J of Separation Science

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Advances in bioanalytical technology have accelerated the analysis of complex protein glycosylation, which is beneficial to understand glycosylation in drug discovery and disease diagnosis. Due to its biological uniqueness in the course of disease occurrence and development, disease‐specific glycosylation requires quantitative characterization of protein glycosylation. We provide a comprehensive review of recent advances in glycosylation analysis, including workflows for glycoprotein digestion, glycopeptide separation and enrichment, and mass spectrometry sequencing. We specifically focus on different strategies for glycopeptide enrichment through physical interaction, chemical oxidation, or metabolic labeling of intact glycopeptides. Recent advances and challenges of O‐glycosylation analysis are presented, and the development of improved enrichment methods combining different proteases to analyze O‐glycosylation is also proposed.

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Section: General Workflow For Analyzing Glycans and Glycopeptidesmentioning

confidence: 99%

Advances in glycopeptide enrichment methods for the analysis of protein glycosylation over the past decade

Zhang

et al. 2022

J of Separation Science

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“…These can be achieved by the release of N-glycans by N-glycosidases and the removal of O-glycans by chemical β-elimination ( Jensen et al., 2012 ; Yang et al., 2017a ), while N-glycosites are determined by hydrophilic interaction liquid chromatography (HILIC) - tandem MS (MS/MS) of intact N-glycopeptides ( Riley et al., 2019 ; Zacharias et al., 2016 ; Xiao et al., 2018 ). Complex O-glycosylation has been successfully studied by O-protease, which can cleave the N-terminus of O-glycosylated Ser or Thr , and EThcD (Electron-transfer and higher-energy collision dissociation) ( Yang et al., 2018b , 2020 ; Malaker et al., 2019 ). In contrast, the linkages of labile sialic acids are differentially protected through ethyl esterification and reductive amination using an amine-containing compound ( Reiding et al., 2014 ; Yang et al., 2017b ).…”

Section: Abnormal Glycosylation Of Saliva and Lung Proteins In Lung Cancermentioning

confidence: 99%

Abnormal sialylation and fucosylation of saliva glycoproteins: Characteristics of lung cancer-specific biomarkers

Gao

Yue

et al. 2022

Current Research in Pharmacology and Drug Discovery

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Dysregulated surface glycoproteins play an important role in tumor cell proliferation and progression. Abnormal glycosylation of these glycoproteins may activate tumor signal transduction and lead to tumor development. The tumor microenvironment alters its molecular composition, some of which regulate protein glycosylation biosynthesis. The glycosylation of saliva proteins in lung cancer patients is different from healthy controls, in which the glycans of cancer patients are highly sialylated and hyperfucosylated. Most studies have shown that O-glycans from cancer are truncated O-glycans, while N-glycans contain fucoses and sialic acids. Because glycosylation analysis is challenging, there are few reports on how glycosylation of saliva proteins is related to the occurrence or progression of lung cancer. In this review, we discussed glycoenzymes involved in protein glycosylation, their changes in tumor microenvironment, potential tumor biomarkers present in body fluids, and abnormal glycosylation of saliva or lung glycoproteins. We further explored the effect of glycosylation changes on tumor signal transduction, and emphasized the role of receptor tyrosine kinases in tumorigenesis and metastasis.

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“… 43 − 45 Complex O-glycosylation has been successfully studied by O-protease (OpeRATOR or StcE), which cleaves the N-terminus of O -glycosylated serine or threonine; O -glycopeptides are usually analyzed by electron-transfer and higher-energy collision dissociation (EThcD) fragmentation. 46 − 48 Conversely, the linkages of labile sialic acids are differentially derivatized by ethyl esterification and reductive amination using amine-containing compounds. 49 , 50 The derivatization of sialic acid on the solid phase not only stabilizes the α-2,3 and α-2,6 linkages sequentially but also facilitates the removal of reagents after the reaction.…”

Section: Introductionmentioning

confidence: 99%

Aberrant Fucosylation of Saliva Glycoprotein Defining Lung Adenocarcinomas Malignancy

Gao

Han

et al. 2022

ACS Omega

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Aberrant glycosylation is a hallmark of cancer found during tumorigenesis and tumor progression. Lung cancer (LC) induced by oncogene mutations has been detected in the patient’s saliva, and saliva glycosylation has been altered. Saliva contains highly glycosylated glycoproteins, the characteristics of which may be related to various diseases. Therefore, elucidating cancer-specific glycosylation in the saliva of healthy, non-cancer, and cancer patients can reveal whether tumor glycosylation has unique characteristics for early diagnosis. In this work, we used a solid-phase chemoenzymatic method to study the glycosylation of saliva glycoproteins in clinical specimens. The results showed that the α1,6-core fucosylation of glycoproteins was increased in cancer patients, whereas α1,2 or α1,3 fucosylation was significantly increased. We further analyzed the expression of fucosyltransferases responsible for α1,2, α1,3, and α1,6 fucosylation. The fucosylation of the saliva of cancer patients is drastically different from that of non-cancer or health controls. These results indicate that the glycoform of saliva fucosylation distinguishes LC from other diseases, and this feature has the potential to diagnose lung adenocarcinoma.

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Improved online LC-MS/MS identification of O-glycosites by EThcD fragmentation, chemoenzymatic reaction, and SPE enrichment

Cited by 15 publications

References 37 publications

Advances in glycopeptide enrichment methods for the analysis of protein glycosylation over the past decade

Advances in glycopeptide enrichment methods for the analysis of protein glycosylation over the past decade

Abnormal sialylation and fucosylation of saliva glycoproteins: Characteristics of lung cancer-specific biomarkers

Aberrant Fucosylation of Saliva Glycoprotein Defining Lung Adenocarcinomas Malignancy

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