Automated glycan assembly of galactosylated xyloglucan oligosaccharides and their recognition by plant cell wall glycan-directed antibodies

Dallabernardina, Pietro; Ruprecht, Colin; Smith, Peter James; Hahn, Michael G.; Urbanowicz, Breeanna R.; Pfrengle, Fabian

doi:10.1039/c7ob02605f

Cited by 29 publications

(34 citation statements)

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“…To systematically map the acceptor specificity of XETs, we chemically synthesized a set of xyloglucan oligosaccharides (XGOs) by automated glycan assembly according to previously published procedures . This set contained oligosaccharides comprising β‐1,4‐linked glucotetraosyl backbones appended with single α‐1,6‐linked xylose substitutions in the +1 position counting from the nonreducing end of the acceptor (XGGG), +2 position (GXGG), and +3 position (GGXG), as well as with two xylose substitutions in the +2 and +3 positions (GXXG) . The oligosaccharides are denoted according to the common nomenclature for xyloglucans (Figure ) .…”

Section: Figuresupporting

confidence: 81%

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Analyzing Xyloglucan Endotransglycosylases by Incorporating Synthetic Oligosaccharides into Plant Cell Walls

et al. 2018

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The plant cell wall is a cellular exoskeleton consisting predominantly of a complex polysaccharide network that defines the shape of cells. During growth, this network can be loosened through the action of xyloglucan endotransglycosylases (XETs), glycoside hydrolases that "cut and paste" xyloglucan polysaccharides through a transglycosylation process. We have analyzed cohorts of XETs in different plant species to evaluate the substrate specificities of xyloglucan acceptors by using a set of synthetic oligosaccharides obtained by automated glycan assembly. The ability of XETs to incorporate the oligosaccharides into polysaccharides printed as microarrays and into stem sections of Arabidopsis thaliana, beans, and peas was assessed. We found that single xylose substitutions are sufficient for transfer, and xylosylation of the terminal glucose residue is not required by XETs, independent of plant species. To obtain information on the potential xylosylation pattern of the natural acceptor of XETs, that is, the nonreducing end of xyloglucan, we further tested the activity of xyloglucan xylosyl transferase (XXT) 2 on the synthetic xyloglucan oligosaccharides. These data shed light on inconsistencies between previous studies towards determining the acceptor substrate specificities of XETs and have important implications for further understanding plant cell wall polysaccharide synthesis and remodeling.

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Section: Figuresupporting

confidence: 81%

“…The syntheses of XGGG, GXGG, and GGXG are described in detail in ref. , and the synthesis of GXXG is described in ref. .…”

Section: Figurementioning

confidence: 99%

Analyzing Xyloglucan Endotransglycosylases by Incorporating Synthetic Oligosaccharides into Plant Cell Walls

et al. 2018

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“…As for xyloglucan and pectin epitopes, three antibodies successfully recognized the modifications between EA-CS and UT-CS including the antibodies CCRC-M88, CCRC-M14 and CCRC-M38. The antibody CCRC-M88 ( Figure 5III, A and D) recognizes galactosylated xyloglucan epitopes (Dallabernardina et al, 2017). The labeling with this antibody was stronger in the phloem and parenchyma cell walls in the control (Figure 5III, A), while, the labeling was left in the phloem area after the pretreatment (Figure 5III, D).…”

Section: Understanding Cell Wall Ultrastructure Using Immunofluorescementioning

confidence: 98%

Effects of Extractive Ammonia Pretreatment on the Ultrastructure and Glycan Composition of Corn Stover

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Zhou

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et al. 2019

Front. Energy Res.

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“…Non-FUC-XyG epitopes were detected by four mAb clusters in SYP61 vesicles ( Figure 3B; Supplemental Figures 2 and 3; Supplemental Data Set 3). In these clusters, some prominent Abs (CCRC-M87, CCRC-M88, CCRC-M93, CCRC-M95, CCRC-M101, and CCRC-M104; Figure 3B, indicated by circles on the left side of the heatmap) bind to galactosylated XyG epitopes (XLXG, XXLG, and XLLG; Dallabernardina et al, 2017). These galactosylated XyG epitopes were also present in the biosynthetic compartment (Golgi/TGN-enriched fraction; Figure 3A; Supplemental Data Set 1A) and cell wall extracts ( Figure 3C; Supplemental Data Set 4).…”

Section: Diverse Xyloglucan Epitopes Are Present In Syp61 Vesiclesmentioning

confidence: 99%

A Hybrid Approach Enabling Large-Scale Glycomic Analysis of Post-Golgi Vesicles Reveals a Transport Route for Polysaccharides

et al. 2019

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The plant endomembrane system facilitates the transport of polysaccharides, associated enzymes, and glycoproteins through its dynamic pathways. Although enzymes involved in cell wall biosynthesis have been identified, little is known about the endomembrane-based transport of glycan components. This is partially attributed to technical challenges in biochemically determining polysaccharide cargo in specific vesicles. Here, we introduce a hybrid approach addressing this limitation. By combining vesicle isolation with a large-scale carbohydrate antibody arraying technique, we charted an initial large-scale map describing the glycome profile of the SYNTAXIN OF PLANTS61 (SYP61) trans-Golgi network compartment in Arabidopsis (Arabidopsis thaliana). A library of antibodies recognizing specific noncellulosic carbohydrate epitopes allowed us to identify a range of diverse glycans, including pectins, xyloglucans (XyGs), and arabinogalactan proteins in isolated vesicles. Changes in XyG-and pectin-specific epitopes in the cell wall of an Arabidopsis SYP61 mutant corroborate our findings. Our data provide evidence that SYP61 vesicles are involved in the transport and deposition of structural polysaccharides and glycoproteins. Adaptation of our methodology can enable studies characterizing the glycome profiles of various vesicle populations in plant and animal systems and their respective roles in glycan transport defined by subcellular markers, developmental stages, or environmental stimuli.

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Automated glycan assembly of galactosylated xyloglucan oligosaccharides and their recognition by plant cell wall glycan-directed antibodies

Cited by 29 publications

References 29 publications

Analyzing Xyloglucan Endotransglycosylases by Incorporating Synthetic Oligosaccharides into Plant Cell Walls

Analyzing Xyloglucan Endotransglycosylases by Incorporating Synthetic Oligosaccharides into Plant Cell Walls

Effects of Extractive Ammonia Pretreatment on the Ultrastructure and Glycan Composition of Corn Stover

A Hybrid Approach Enabling Large-Scale Glycomic Analysis of Post-Golgi Vesicles Reveals a Transport Route for Polysaccharides

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