Contributions to Arabinogalactan Protein Analysis

Castilleux, Romain; Ropitaux, Marc; Manasfi, Youssef; Bernard, Sophie; Vicré-Gibouin, Maïté; Driouich, Azeddine

doi:10.1007/978-1-0716-0621-6_22

Cited by 6 publications

(3 citation statements)

References 33 publications

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“…Detection of AGPs on agarose gel was performed according to Castilleux et al (2020) . Water-extracted AGPs from 15 mg pooled samples were loaded onto a 1% (w/v) agarose gel containing 90 mM Tris base pH 8.3 with HCl, 90 mM boric acid, and 2 mM Na 2 EDTA (H 2 O) 2 , and run at 100 V for 1 h. β-D-glucosyl Yariv was synthesized in house according to the protocol by Yariv et al (1962) .…”

Section: Methodsmentioning

confidence: 99%

The Spatio-Temporal Distribution of Cell Wall-Associated Glycoproteins During Wood Formation in Populus

et al. 2020

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Plant cell wall associated hydroxyproline-rich glycoproteins (HRGPs) are involved in several aspects of plant growth and development, including wood formation in trees. HRGPs such as arabinogalactan-proteins (AGPs), extensins (EXTs), and proline rich proteins (PRPs) are important for the development and architecture of plant cell walls. Analysis of publicly available gene expression data revealed that many HRGP encoding genes show tight spatio-temporal expression patterns in the developing wood of Populus that are indicative of specific functions during wood formation. Similar results were obtained for the expression of glycosyl transferases putatively involved in HRGP glycosylation. In situ immunolabelling of transverse wood sections using AGP and EXT antibodies revealed the cell type specificity of different epitopes. In mature wood AGP epitopes were located in xylem ray cell walls, whereas EXT epitopes were specifically observed between neighboring xylem vessels, and on the ray cell side of the vessel walls, likely in association with pits. Molecular mass and glycan analysis of AGPs and EXTs in phloem/cambium, developing xylem, and mature xylem revealed clear differences in glycan structures and size between the tissues. Separation of AGPs by agarose gel electrophoresis and staining with β-D-glucosyl Yariv confirmed the presence of different AGP populations in phloem/cambium and xylem. These results reveal the diverse changes in HRGP-related processes that occur during wood formation at the gene expression and HRGP glycan biosynthesis levels, and relate HRGPs and glycosylation processes to the developmental processes of wood formation.

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Section: Methodsmentioning

confidence: 99%

The Spatio-Temporal Distribution of Cell Wall-Associated Glycoproteins During Wood Formation in Populus

et al. 2020

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“…For rapid detection and semi-quantification of AGPs, the radial gel diffusion assay using Yariv specific reactivity was carried out as in Holst and Clarke [27] and Castilleux [28]. The samples, i.e.…”

Section: Yariv Reagent Reactivity -Radial Gel Diffusion Assaymentioning

confidence: 99%

Working towards arabinogalactan proteins (AGPs) from fruit: carbohydrate composition and impact on fungal growth

et al. 2022

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Background Arabinogalactan proteins (AGPs) are extracellular matrix constituents involved in plant response to fungal infection. The aim of the current study was to investigate the antifungal effect of AGPs ex situ and to determine the structural features of AGPs that may have an influence on this activity. The features of AGPs isolated from fruit were investigated with molecular tools based on specific monoclonal antibodies recognizing carbohydrate AGP epitopes. The Antifungal (well-diffusion) Susceptibility Test and the Agar Invasion Test were used to assess the impact of AGPs on Penicillium notatum culture. Results The results definitely ruled out the influence of AGPs on fungal growth. The immunochemical analyses revealed that AGPs consist mainly of carbohydrate chains composed of β-linked glucuronosyl residues recognized by LM2 and GlcA-β(1 → 3)-GalA-α(1 → 2) Rha recognized by JIM13, which do not have the same functional properties outside the plant cell in in vitro experimental conditions. Conclusions The action of a single cell wall component does not elicit any influence ex situ. The extensive accumulation of glycan chains of AGPs in infected tissue as a result of a complex mechanism occurring in the cell wall emphasizes the importance of dependencies between particular components of the extracellular matrix in response to fungal attack.

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“…From the experimental point of view, immunolabeling with antibodies or the use of β-Yariv reagents is the most commonly used (Nguema-Ona et al, 2012). Compared to the monoclonal antibodies, the β-Yariv reagents are able (in addition to the visualization of AGPs) also to perturb their function, which is widely exploited in AGP functional studies (Willats and Knox, 1996;Tang et al, 2006;Nguema-Ona et al, 2007;Yu and Zhao, 2012;Olmos et al, 2017;Su and Higashiyama, 2018;Castilleux et al, 2020).…”

Section: Introductionmentioning

confidence: 99%

Structural Basis of the Function of Yariv Reagent—An Important Tool to Study Arabinogalactan Proteins

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Pavlů

Hancharyk

et al. 2021

Front. Mol. Biosci.

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Arabinogalactan proteins are very abundant, heavily glycosylated plant cell wall proteins. They are intensively studied because of their crucial role in plant development as well as their function in plant defence. Research of these biomacromolecules is complicated by the lack of tools for their analysis and characterisation due to their extreme heterogeneity. One of the few available tools for detection, isolation, characterisation, and functional studies of arabinogalactan proteins is Yariv reagents. Yariv reagent is a synthetic aromatic glycoconjugate originally prepared as an antigen for immunization. Later, it was found that this compound can precipitate arabinogalactan proteins, namely, their ß-D-(1→3)-galactan structures. Even though this compound has been intensively used for decades, the structural basis of arabinogalactan protein precipitation by Yariv is not known. Multiple biophysical studies have been published, but none of them attempted to elucidate the three-dimensional structure of the Yariv-galactan complex. Here we use a series of molecular dynamics simulations of systems containing one or multiple molecules of ß-D-galactosyl Yariv reagent with or without oligo ß-D-(1→3)-galactan to predict the structure of the complex. According to our model of Yariv-galactan complexes, Yariv reagent forms stacked oligomers stabilized by π-π and CH/π interactions. These oligomers may contain irregularities. Galactan structures crosslink these Yariv oligomers. The results were compared with studies in literature.

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Contributions to Arabinogalactan Protein Analysis

Cited by 6 publications

References 33 publications

The Spatio-Temporal Distribution of Cell Wall-Associated Glycoproteins During Wood Formation in Populus

The Spatio-Temporal Distribution of Cell Wall-Associated Glycoproteins During Wood Formation in Populus

Working towards arabinogalactan proteins (AGPs) from fruit: carbohydrate composition and impact on fungal growth

Structural Basis of the Function of Yariv Reagent—An Important Tool to Study Arabinogalactan Proteins

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