Automated Synthesis of Arabinoxylan‐Oligosaccharides Enables Characterization of Antibodies that Recognize Plant Cell Wall Glycans

Schmidt, Deborah; Schuhmacher, Frank; Geissner, Andreas; Seeberger, Peter H.; Pfrengle, Fabian

doi:10.1002/chem.201500065

Cited by 94 publications

(100 citation statements)

References 41 publications

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“…Welldefined, single-compound oligosaccharides provide the highest resolution possible when characterizing protein-carbohydrate interactions, provided that the binding interactions are sufficiently strong to be detected. This is especially true when a range of structurally related but distinct glycans are available (for examples, see Clausen et al, 2003;Andersen et al, 2016aAndersen et al, , 2016bSchmidt et al, 2015). In the case of LM26 discussed here, it would have been close to impossible to identify the epitope without access to the appropriate and defined linear and branched galactans.…”

Section: Discussion Synthetic Glycan Arrays As a Discovery Platformmentioning

confidence: 97%

Branched Pectic Galactan in Phloem-Sieve-Element Cell Walls: Implications for Cell Mechanics

et al. 2017

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A major question in plant biology concerns the specification and functional differentiation of cell types. This is in the context of constraints imposed by networks of cell walls that both adhere cells and contribute to the form and function of developing organs. Here, we report the identification of a glycan epitope that is specific to phloem sieve element cell walls in several systems. A monoclonal antibody, designated LM26, binds to the cell wall of phloem sieve elements in stems of Arabidopsis (Arabidopsis thaliana), Miscanthus x giganteus, and notably sugar beet (Beta vulgaris) roots where phloem identification is an important factor for the study of phloem unloading of Suc. Using microarrays of synthetic oligosaccharides, the LM26 epitope has been identified as a b-1,6-galactosyl substitution of b-1,4-galactan requiring more than three backbone residues for optimized recognition. This branched galactan structure has previously been identified in garlic (Allium sativum) bulbs in which the LM26 epitope is widespread throughout most cell walls including those of phloem cells. Garlic bulb cell wall material has been used to confirm the association of the LM26 epitope with cell wall pectic rhamnogalacturonan-I polysaccharides. In the phloem tissues of grass stems, the LM26 epitope has a complementary pattern to that of the LM5 linear b-1,4-galactan epitope, which is detected only in companion cell walls. Mechanical probing of transverse sections of M. x giganteus stems and leaves by atomic force microscopy indicates that phloem sieve element cell walls have a lower indentation modulus (indicative of higher elasticity) than companion cell walls.

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Section: Discussion Synthetic Glycan Arrays As a Discovery Platformmentioning

confidence: 97%

Branched Pectic Galactan in Phloem-Sieve-Element Cell Walls: Implications for Cell Mechanics

et al. 2017

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“…In accordance with the biochemical composition analysis (Table III), mucilage displayed less intense CCRC-M139 signals compared with RG I labeled by CCRC-M36. Although its precise epitope structure remains to be confirmed, CCRC-M139 can bind unbranched xylan with degrees of polymerization of at least 6 (Schmidt et al, 2015). CCRC-M139 epitopes were only detected at the outer edge of wild-type adherent mucilage (Fig.…”

Section: Highly Branched Xylan Attaches Mucilage Pectin To Cellulosicmentioning

confidence: 98%

Highly Branched Xylan Made by IRX14 and MUCI21 Links Mucilage to Arabidopsis Seeds

et al. 2015

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ORCID IDs: 0000-0001-9105-014X (C.V.); 0000-0003-4576-6774 (M.H.-W.S.).All cells of terrestrial plants are fortified by walls composed of crystalline cellulose microfibrils and a variety of matrix polymers. Xylans are the second most abundant type of polysaccharides on Earth. Previous studies of Arabidopsis (Arabidopsis thaliana) irregular xylem (irx) mutants, with collapsed xylem vessels and dwarfed stature, highlighted the importance of this cell wall component and revealed multiple players required for its synthesis. Nevertheless, xylan elongation and substitution are complex processes that remain poorly understood. Recently, seed coat epidermal cells were shown to provide an excellent system for deciphering hemicellulose production. Using a coexpression and sequence-based strategy, we predicted several MUCILAGE-RELATED (MUCI) genes that encode glycosyltransferases (GTs) involved in the production of xylan. We now show that MUCI21, a member of an uncharacterized clade of the GT61 family, and IRX14 (GT43 protein) are essential for the synthesis of highly branched xylan in seed coat epidermal cells. Our results reveal that xylan is the most abundant xylose-rich component in Arabidopsis seed mucilage and is required to maintain its architecture. Characterization of muci21 and irx14 single and double mutants indicates that MUCI21 is a Golgi-localized protein that likely facilitates the addition of xylose residues directly to the xylan backbone. These unique branches seem to be necessary for pectin attachment to the seed surface, while the xylan backbone maintains cellulose distribution. Evaluation of muci21 and irx14 alongside mutants that disrupt other wall components suggests that mucilage adherence is maintained by complex interactions between several polymers: cellulose, xylans, pectins, and glycoproteins.

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“…The de novo synthesis of various oligosaccharides has enabled the characterization of xylan-directed mAbs (Schmidt et al, 2015). While microarrays with plant polysaccharides (Moller et al, 2008) or oligosaccharides (Pedersen et al, 2012) have been probed previously with mAbs (Pattathil et al, 2010), arrays of synthetic glycans have an exceptional purity and facilitate the precise mapping of the epitopes recognized by molecular probes.…”

Section: Xylan Dynamics In Secondary Cell Wallsmentioning

confidence: 99%

Monitoring Polysaccharide Dynamics in the Plant Cell Wall

2018

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Automated Synthesis of Arabinoxylan‐Oligosaccharides Enables Characterization of Antibodies that Recognize Plant Cell Wall Glycans

Cited by 94 publications

References 41 publications

Branched Pectic Galactan in Phloem-Sieve-Element Cell Walls: Implications for Cell Mechanics

Branched Pectic Galactan in Phloem-Sieve-Element Cell Walls: Implications for Cell Mechanics

Highly Branched Xylan Made by IRX14 and MUCI21 Links Mucilage to Arabidopsis Seeds

Monitoring Polysaccharide Dynamics in the Plant Cell Wall

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