Structural Glycomic Analyses at High Sensitivity: A Decade of Progress

Alley, William R.; Novotný, Miloš V.

doi:10.1146/annurev-anchem-062012-092609

Cited by 60 publications

(51 citation statements)

References 146 publications

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“…For this group of glycans, eight distinct core structures have been discovered (illustrated in Figure 2). It is noteworthy that some of these structures (e.g., Core 1 and Core 8) share the same glycan compositions but differ in their linkage positions between adjacent monosaccharide units (15,18). Additional carbohydrates are often added to these core structures through enzyme-controlled reactions to generate O-linked glycans of diverse branching and elongation patterns.…”

Section: O-linked Glycosylationmentioning

confidence: 99%

Carbohydrates on Proteins: Site-Specific Glycosylation Analysis by Mass Spectrometry

Zhu

Desaire²

2015

Annual Rev. Anal. Chem.

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Glycosylation on proteins adds complexity and versatility to these biologically vital macromolecules. To unveil the structure-function relationship of glycoproteins, glycopeptide-centric analysis using mass spectrometry (MS) has become a method of choice because the glycan is preserved on the glycosylation site and site-specific glycosylation profiles of proteins can be readily determined. However, glycopeptide analysis is still challenging given that glycopeptides are usually low in abundance and relatively difficult to detect and the resulting data require expertise to analyze. Viewing the urgent need to address these challenges, emerging methods and techniques are being developed with the goal of analyzing glycopeptides in a sensitive, comprehensive, and high-throughput manner. In this review, we discuss recent advances in glycoprotein and glycopeptide analysis, with topics covering sample preparation, analytical separation, MS and tandem MS techniques, as well as data interpretation and automation.

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Section: O-linked Glycosylationmentioning

confidence: 99%

Carbohydrates on Proteins: Site-Specific Glycosylation Analysis by Mass Spectrometry

Zhu

Desaire²

2015

Annual Rev. Anal. Chem.

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“…For the assignment of glycan structures tandem mass spectrometry (MS/MS) is a particularly powerful tool. Here, selected precursor ions are fragmented into product ions, usually by the collision with a gas, and the fragmentation pattern employed to gain additional insight into the analyte structure [53]. …”

Section: Ev Glycomicsmentioning

confidence: 99%

Glycosylation of extracellular vesicles: current knowledge, tools and clinical perspectives

Williams

Royo

Aizpurua‐Olaizola

et al. 2018

J of Extracellular Vesicle

209

163

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It is now acknowledged that extracellular vesicles (EVs) are important effectors in a vast number of biological processes through intercellular transfer of biomolecules. Increasing research efforts in the EV field have yielded an appreciation for the potential role of glycans in EV function. Indeed, recent reports show that the presence of glycoconjugates is involved in EV biogenesis, in cellular recognition and in the efficient uptake of EVs by recipient cells. It is clear that a full understanding of EV biology will require researchers to focus also on EV glycosylation through glycomics approaches. This review outlines the major glycomics techniques that have been applied to EVs in the context of the recent findings. Beyond understanding the mechanisms by which EVs mediate their physiological functions, glycosylation also provides opportunities by which to engineer EVs for therapeutic and diagnostic purposes. Studies characterising the glycan composition of EVs have highlighted glycome changes in various disease states, thus indicating potential for EV glycans as diagnostic markers. Meanwhile, glycans have been targeted as molecular handles for affinity-based isolation in both research and clinical contexts. An overview of current strategies to exploit EV glycosylation and a discussion of the implications of recent findings for the burgeoning EV industry follows the below review of glycomics and its application to EV biology.

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“…Recent years have seen an increase in the possibilities for glycomic analyses of high sensitivity (30) ; a primary tool is mass spectrometry, especially of LC-ESI-MS and MALDI-TOF-MS of native and permethylated N-glycans. As in a number of our previouslypublished studies, we have opted here for an off-line LC-MALDI-TOF MS approach in order to separate the N-glycans into fractions of lower complexity and facilitate their analysis by chemical and enzymatic treatments on a small scale.…”

Section: Analysis and Chromatography Of N-glycans Of Lower Eukaryotesmentioning

confidence: 99%

N‐glycomic profiling of a glucosidase II mutant of Dictyostelium discoideum by ‘‘off‐line’’ liquid chromatography and mass spectrometry

et al. 2014

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In this study, we have performed the first mass spectrometric analysis of N-glycans of the M31 mutant strain of the cellular slime mould Dictyostelium discoideum, previously shown to have a defect in glucosidase II. Together with glucosidase I, this enzyme mediates part of the initial processing of N-glycans; defects in either glucosidase are associated with human diseases and result in an accumulation of incorrectly-processed oligosaccharides which are not, or only poor, substrates for a range of downstream enzymes. To examine the effect of the glucosidase II mutation in Dictyostelium, we employed off-line LC-MALDI-TOF-MS in combination with chemical and enzymatic treatments and MS/MS to analyse the neutral and anionic N-glycans of the mutant as compared to the wild-type. The major neutral species were, as expected, of the composition Hex 10-11 HexNAc 2-3 with one or two terminal glucose residues. Consistent with the block in processing of neutral N-glycans caused by the absence of glucosidase II, fucose was apparently absent from the N-glycans and bisecting N-acetylglucosamine was rare. The major anionic oligosaccharides were sulphated and/or methylphosphorylated forms of Hex 8-11 HexNAc 2-3 , many of which surprisingly lacked glucose residues entirely. As anionic Nglycans are considered to be mostly associated with lysosomal enzymes in Dictyostelium, we hypothesise that glycosidases present in the acidic compartments may act on the oligosaccharides attached to such slime mould proteins. Furthermore, our chosen analytical approach enabled us, via observation of diagnostic negative-mode MS/MS fragments, to determine the fine structure of the methylphosphorylated and sulphated N-glycans of the M31 glucosidase mutant in their native state.

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Structural Glycomic Analyses at High Sensitivity: A Decade of Progress

Cited by 60 publications

References 146 publications

Carbohydrates on Proteins: Site-Specific Glycosylation Analysis by Mass Spectrometry

Carbohydrates on Proteins: Site-Specific Glycosylation Analysis by Mass Spectrometry

Glycosylation of extracellular vesicles: current knowledge, tools and clinical perspectives

N‐glycomic profiling of a glucosidase II mutant of Dictyostelium discoideum by ‘‘off‐line’’ liquid chromatography and mass spectrometry

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