Comprehensive structural glycomic characterization of the glycocalyxes of cells and tissues

Li, Qiongyu; Xie, Yixuan; Wong, Maurice; Barboza, Mariana; Lebrilla, Carlito B.

doi:10.1038/s41596-020-0350-4

Cited by 66 publications

(86 citation statements)

References 84 publications

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“…PGC columns can separate closely related unlabeled glycan isomers that differ only in arm position and sialyl linkages. PGC chips equipped for nanoLC-MS have also been developed for analysis of small amounts of glycan samples 26 . However, evaluating the ability of PGC to separate glycan isomers from various kinds of tissue samples will require accumulation of additional data, as it remains ambiguous in terms of discriminations of branching patterns of multiantennary structures or the number of LacNAc repeats.…”

mentioning

confidence: 99%

Toward robust N-glycomics of various tissue samples that may contain glycans with unknown or unexpected structures

Suzuki

Abé

Hanzawa

et al. 2021

Sci Rep

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Glycans in tissues are structurally diverse and usually include a large number of isomers that cannot be easily distinguished by mass spectrometry (MS). To address this issue, we developed a combined method that can efficiently separate and identify glycan isomers. First, we separated 2-aminopyridine (PA)-derivatized N-glycans from chicken colon by reversed-phase liquid chromatography (LC) and directly analyzed them by electrospray ionization (ESI)-MS and MS/MS to obtain an overview of the structural features of tissue glycans. Next, we deduced the structures of isomers based on their elution positions, full MS, and MS/MS data, before or after digestions with several exoglycosidases. In this method, the elution position differed greatly depending on the core structure and branching pattern, allowing multiantennary N-glycan structures to be easily distinguished. To further determine linkages of branch sequences, we modified PA-N-glycans with sialic acid linkage-specific alkylamidation and/or permethylation, and analyzed the products by LC–MS and multistage MS. We determined the relative abundances of core structures, branching patterns, and branch sequences of N-glycans from chicken colon, and confirmed presence of characteristic branch sequences such as Lex, sialyl Lex, sulfated LacNAc, LacNAc repeat, and LacdiNAc. The results demonstrated that our method is useful for comparing N-glycomes among various tissue samples.

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mentioning

confidence: 99%

Toward robust N-glycomics of various tissue samples that may contain glycans with unknown or unexpected structures

Suzuki

Abé

Hanzawa

et al. 2021

Sci Rep

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“…Currently, no singular strategy is available that enables the simultaneous identification of peptide sequence and full glycan structure characterization of glycoproteins [52,61]. Rather, multiple complementary strategies are required to fully elucidate the glycan structures present on specific sites within a protein (Fig.…”

Section: Strategies To Assess Protein Glycosylationmentioning

confidence: 99%

Importance of evaluating protein glycosylation in pluripotent stem cell-derived cardiomyocytes for research and clinical applications

Kelly

Albahrani

Castro

et al. 2021

Pflugers Arch - Eur J Physiol

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Proper protein glycosylation is critical to normal cardiomyocyte physiology. Aberrant glycosylation can alter protein localization, structure, drug interactions, and cellular function. The in vitro differentiation of human pluripotent stem cells into cardiomyocytes (hPSC-CM) has become increasingly important to the study of protein function and to the fields of cardiac disease modeling, drug testing, drug discovery, and regenerative medicine. Here, we offer our perspective on the importance of protein glycosylation in hPSC-CM. Protein glycosylation is dynamic in hPSC-CM, but the timing and extent of glycosylation are still poorly defined. We provide new data highlighting how observed changes in hPSC-CM glycosylation may be caused by underlying differences in the protein or transcript abundance of enzymes involved in building and trimming the glycan structures or glycoprotein gene products. We also provide evidence that alternative splicing results in altered sites of glycosylation within the protein sequence. Our findings suggest the need to precisely define protein glycosylation events that may have a critical impact on the function and maturation state of hPSC-CM. Finally, we provide an overview of analytical strategies available for studying protein glycosylation and identify opportunities for the development of new bioinformatic approaches to integrate diverse protein glycosylation data types. We predict that these tools will promote the accurate assessment of protein glycosylation in future studies of hPSC-CM that will ultimately be of significant experimental and clinical benefit.

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“…In this work, we present a sensitive CSF glycan analytical strategy that used as little as 15 μL of starting material; as far as we know, there is no other analytical application using such a small sample volume with reproducible results. The sample preparation was complemented with a SPE-C18 (solid phase extraction) glycan purification to avoid undesirable matrix components [34]. This was followed by permethylation that enhanced the glycan hydrophobicity, thereby increasing ionization efficiency and rendering them amenable to positive ionization.…”

Section: Resultsmentioning

confidence: 99%

N-Glycomics of Cerebrospinal Fluid: Method Comparison

2021

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Cerebrospinal fluid (CSF) contains valuable biological and neurological information. However, its glycomics analysis is hampered due to the low amount of protein in the biofluid, as has been demonstrated by other glycomics studies using a substantial amount of CSF. In this work, we investigated different N-glycan sample preparation approaches to develop a more sensitive method. These methods, one with an increased amount of buffer solution during the N-glycan release step with a lower amount of sample volume and the other with Filter-Aided N-Glycan Separation (FANGS), were compared with recent work to demonstrate their effectiveness. It was demonstrated that an increased amount of buffer solution showed higher intensity in comparison to the previously published method and FANGS. This suggested that digestion efficiency during the N-glycan release step was not in an optimal condition from the previously published method, and that there is a substantial loss of sample with FANGS when preparing N-glycans from CSF.

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Comprehensive structural glycomic characterization of the glycocalyxes of cells and tissues

Cited by 66 publications

References 84 publications

Toward robust N-glycomics of various tissue samples that may contain glycans with unknown or unexpected structures

Toward robust N-glycomics of various tissue samples that may contain glycans with unknown or unexpected structures

Importance of evaluating protein glycosylation in pluripotent stem cell-derived cardiomyocytes for research and clinical applications

N-Glycomics of Cerebrospinal Fluid: Method Comparison

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