Glycan Engineering for Cell and Developmental Biology

Griffin, Matthew E.; Hsieh‐Wilson, Linda C.

doi:10.1016/j.chembiol.2015.12.007

Cited by 78 publications

(88 citation statements)

References 99 publications

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“…In the past decade, metabolic labeling using clickable carbohydrate (Baskin, J.M., Dehnert, K.W., et al 2010, Codelli, J.A., Baskin, J.M., et al 2008, Hsu, T.L., Hanson, S.R., et al 2007, Kizuka, Y., Funayama, S., et al 2016 has revolutionized glycan imaging, as it eliminates the problems associated with low binding affinity via covalent bonding. Subsequently, enzymatic glycan labeling using clickable sugars have been reported (Griffin, M.E. and Hsieh-Wilson, L.C.…”

Section: Introductionmentioning

confidence: 99%

Imaging specific cellular glycan structures using glycosyltransferases via click chemistry

Person

Anderson

et al. 2017

Preprint

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Heparan sulfate (HS) is a polysaccharide fundamentally important for biologically activities. T/Tn antigens are universal carbohydrate cancer markers. Here, we report the specific imaging of these carbohydrates using a mesenchymal stem cell line and human umbilical vein endothelial cells (HUVEC). The staining specificities were demonstrated by comparing imaging of different glycans and validated by either removal of target glycans, which results in loss of signal, or installation of target glycans, which results in gain of signal. As controls, representative key glycans including O-GlcNAc, lactosaminyl glycans and hyaluronan were also imaged. HS staining revealed novel architectural features of the extracellular matrix (ECM) of HUVEC cells.Results from T/Tn antigen staining suggest that O-GalNAcylation is a rate-limiting step for Oglycan synthesis. Overall, these highly specific approaches for HS and T/Tn antigen imaging should greatly facilitate the detection and functional characterization of these biologically important glycans.

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Section: Introductionmentioning

confidence: 99%

Imaging specific cellular glycan structures using glycosyltransferases via click chemistry

Person

Anderson

et al. 2017

Preprint

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“…Glycomics is the systematic study of glycan structures, including monosaccharide compositions, glycan sequence, branching pattern and linkage specifications, which are essential to the functions of glycoconjugates [11–14]. Nevertheless, glycomics analysis is inherently challenging due to their structure complexity and heterogeneity, low natural abundance and the lack of amplification methods [5,14,15].…”

Section: Introductionmentioning

confidence: 99%

Sensitive and robust MALDI-TOF-MS glycomics analysis enabled by Girard's reagent T on-target derivatization (GTOD) of reducing glycans

Zhang

Wang

Jin

et al. 2019

Analytica Chimica Acta

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Sensitive glycomics analysis by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) is of great importance but significantly hampered by their low ionization efficiency and labile sialic acid moieties. Chemical derivatization offers a viable way to improve both the ionization efficiency and analytical sensitivity of the glycans in MS analysis by altering their hydrophobicity or charge property. Here we employed Girard's reagent T (GT) for on-target derivatization (GTOD) of reducing glycan under mild acid condition to form stable hydrazones at room temperature, allowing rapid and sensitive identification of neutral and sialylated glycans in positive-ion mode as only permanently positive charged molecular ions without multiple ion adducts by MALDI-TOF-MS. The MS signal intensities of lactose, sialylated N-glycans derived from bovine fetuin and neutral N-glycans derived from RNaseB and ovalbumin were boosted by 7.44, 9.13, 12.96 and 13.47 folds on average (n = 3), respectively. More importantly, after GTOD strategy, unwanted desialylation of sialylated glycans during MS was suppressed. The detection limit of the assay is desirable since the nanogram of N-glycans derived from 0.16 μg ovalbumin could be detected. The assay demonstrated good stability (RSD≤2.95%, within 10 days), reliable reproducibility (RSD = 2.96%, n = 7) and a desirable linear dynamic range from 78 nmol/mL to 10 μmol/mL. The strategy has been successfully applied to MS analysis of reducing glycans from human milks, neutral and sialylated O-, N-glycans from glycoproteins, and reducing glycans derived from glycosphingolipids, presenting neater [M]+ signals which allow detection of more low-abundance glycans and assignation of Neu5Ac vs. Neu5Gc or fucose vs. hexose in glycans due to the absence of the ambiguous interpretation from multiple peaks (ion adducts [M+Na]+ and [M+K]+). Moreover, the GTOD assay prevents desialylation during MALDI-TOF-MS profiling and enables distinct linkage-specific characterization of terminal sialic acids of N-glycans derived from human serum protein when combines with an esterification.

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“…[3] In many instances the role of specific glycans in these processes remains obscure despite the new advances in gene editing [4] and metabolic oligosaccharide engineering (MOE). [5] MOE remodels glycan structures by supplementing the growth media with carbohydrate analogues that get incorporated into glycans by the endogenous biosynthetic machinery. [6] To our knowledge there has been only one report from our previous study where MOE was employed to control a specific signaling pathway, in which we discovered that enhanced fucosylation on N-glycans inhibits Wnt signaling.…”

mentioning

confidence: 99%

Modulating Cell‐Surface Receptor Signaling and Ion Channel Functions by In Situ Glycan Editing

Jiang

López-Aguilar

et al. 2018

Angew Chem Int Ed

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Glycans anchored on cell-surface receptors are active modulators of receptor signaling. A strategy is presented that enforces transient changes to cell-surface glycosylation patterns to tune receptor signaling. This approach, termed in situ glycan editing, exploits recombinant glycosyltransferases to incorporate monosaccharides with linkage specificity onto receptors in situ. α2,3-linked sialic acid or α1,3-linked fucose added in situ suppresses signaling through epidermal growth factor receptor and fibroblast growth factor receptor. We also applied the same strategy to regulate the electrical signaling of a potassium ion channel–human ether-à-go-go-related gene channel. Compared to gene editing, no long-term perturbations are introduced to the treated cells. In situ glycan editing therefore offers a promising approach for studying the dynamic role of specific glycans in membrane receptor signaling and ion channel functions.

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Glycan Engineering for Cell and Developmental Biology

Cited by 78 publications

References 99 publications

Imaging specific cellular glycan structures using glycosyltransferases via click chemistry

Imaging specific cellular glycan structures using glycosyltransferases via click chemistry

Sensitive and robust MALDI-TOF-MS glycomics analysis enabled by Girard's reagent T on-target derivatization (GTOD) of reducing glycans

Modulating Cell‐Surface Receptor Signaling and Ion Channel Functions by In Situ Glycan Editing

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