Large-scale identification of target proteins of a glycosyltransferase isozyme by Lectin-IGOT-LC/MS, an LC/MS-based glycoproteomic approach

Sugahara, Daisuke; Kaji, Hiroyuki; Sugihara, Kazushi; Asano, Masahide; Narimatsu, Hisashi

doi:10.1038/srep00680

Cited by 22 publications

(20 citation statements)

References 40 publications

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“…N-glycosylated peptides were labeled specifically with 18 O using the IGOT reaction as described previously [42]. The HILIC-purified glycopeptides were concentrated.…”

Section: N-glycanase F-mediated 18 O-labeling Of Glycopeptidesmentioning

confidence: 99%

Large‐scale identification of secretome glycoproteins recognized by Wisteria floribunda agglutinin: A glycoproteomic approach to biomarker discovery

et al. 2015

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One of the promising approaches to the development of cancer diagnostic systems is quantification of a specific protein carrying cancerous glycans. Potential utility of Wisteria floribunda agglutinin (WFA) for such assays has been suggested for several cancer types. To develop such diagnostic systems, identification of WFA-recognized glycoproteins is essential. Here, we successfully identified 504 WFA-recognized glycoproteins from the secretome of HEK293T cells. Most of the identified proteins were likely soluble or single-pass transmembrane proteins, which may serve as specific proteins for the diagnosis using biological fluids. Our method may help to discover marker glycoproteins for various cancers generating WFA-recognized glycans.

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“…N-glycosylated peptides were labeled specifically with 18 O using the IGOT reaction as described previously [42]. The HILIC-purified glycopeptides were concentrated.…”

Section: N-glycanase F-mediated 18 O-labeling Of Glycopeptidesmentioning

confidence: 99%

Large‐scale identification of secretome glycoproteins recognized by Wisteria floribunda agglutinin: A glycoproteomic approach to biomarker discovery

et al. 2015

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“…Currently, few systematic studies of glycans in cancer exist to sort out these roles, none related to melanoma. In general, glycomic studies on cancer to date fall short of pinpointing enzymes responsible for the biosynthesis of observed aberrant glycan motifs and/or their relevant glycoprotein targets (Sugahara et al, 2012). Glycosylation remains an understudied portion of cancer biology due to issues including difficulties in glycan analysis and complex biosynthetic pathways (Agre et al, 2016; Rakus and Mahal, 2011).…”

Section: Introductionmentioning

confidence: 99%

A Systems Biology Approach Identifies FUT8 as a Driver of Melanoma Metastasis

et al. 2017

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SUMMARY Association of aberrant glycosylation with melanoma progression is based mainly on analyses of cell lines. Here we present a systems-based study of glycomic changes and corresponding enzymes associated with melanoma metastasis in patient samples. Upregulation of core fucosylation (FUT8) and downregulation of α-1,2 fucosylation (FUT1, FUT2) were identified as features of metastatic melanoma. Using both in vitro and in vivo studies, we demonstrate FUT8 is a driver of melanoma metastasis which, when silenced, suppresses invasion and tumor dissemination. Glycoprotein targets of FUT8 were enriched in cell migration proteins including the adhesion molecule L1CAM. Core fucosylation impacted L1CAM cleavage and the ability of L1CAM to support melanoma invasion. FUT8 and its targets represent therapeutic targets in melanoma metastasis.

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“…However it also has the disadvantage of poor specificity. To improve the enrichment specificity, lectin and HILIC enrichment methods were reported to be applied sequentially 25 26 . The HC method, on the other hand, has the advantage of capturing glycopeptides with high specificity.…”

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confidence: 99%

A peptide N-terminal protection strategy for comprehensive glycoproteome analysis using hydrazide chemistry based method

Huang

Qin

Sun

et al. 2015

Sci Rep

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Enrichment of glycopeptides by hydrazide chemistry (HC) is a popular method for glycoproteomics analysis. However, possible side reactions of peptide backbones during the glycan oxidation in this method have not been comprehensively studied. Here, we developed a proteomics approach to locate such side reactions and found several types of the side reactions that could seriously compromise the performance of glycoproteomics analysis. Particularly, the HC method failed to identify N-terminal Ser/Thr glycopeptides because the oxidation of vicinal amino alcohol on these peptides generates aldehyde groups and after they are covalently coupled to HC beads, these peptides cannot be released by PNGase F for identification. To overcome this drawback, we apply a peptide N-terminal protection strategy in which primary amine groups on peptides are chemically blocked via dimethyl labeling, thus the vicinal amino alcohols on peptide N-termini are eliminated. Our results showed that this strategy successfully prevented the oxidation of peptide N-termini and significantly improved the coverage of glycoproteome.

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Large-scale identification of target proteins of a glycosyltransferase isozyme by Lectin-IGOT-LC/MS, an LC/MS-based glycoproteomic approach

Cited by 22 publications

References 40 publications

Large‐scale identification of secretome glycoproteins recognized by Wisteria floribunda agglutinin: A glycoproteomic approach to biomarker discovery

Large‐scale identification of secretome glycoproteins recognized by Wisteria floribunda agglutinin: A glycoproteomic approach to biomarker discovery

A Systems Biology Approach Identifies FUT8 as a Driver of Melanoma Metastasis

A peptide N-terminal protection strategy for comprehensive glycoproteome analysis using hydrazide chemistry based method

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