Polysaccharide Analysis Using Carbohydrate Gel Electrophoresis: A Method to Study Plant Cell Wall Polysaccharides and Polysaccharide Hydrolases

Goubet, Florence; Jackson, Peter; Deery, Michael J.; Dupree, Paul

doi:10.1006/abio.2001.5444

Cited by 167 publications

(183 citation statements)

References 26 publications

Supporting

Mentioning

179

Contrasting

Order By: Relevance

“…Carbohydrate Gel Electrophoresis-Polysaccharide analysis using carbohydrate gel electrophoresis (PACE) was performed as described (40). Briefly, dry glucans were treated with equal volumes of both, 0.2 M 8-aminonaphtalene-1,3,6-trisulfonic acid (ANTS, Molecular Probes, MoBiTec), prepared in acetic acid/water (3/17, v/v), and 1 M NaCNBH 3 (in Me 2 SO) at 37°C overnight.…”

Section: Methodsmentioning

confidence: 99%

An Ancient Enzyme Domain Hidden in the Putative β-Glucan Elicitor Receptor of Soybean May Play an Active Part in the Perception of Pathogen-associated Molecular Patterns during Broad Host Resistance

Fliegmann

Mithöfer

Wanner

et al. 2004

Journal of Biological Chemistry

148

115

View full text Add to dashboard Cite

A successful defense against potential pathogens requires that a host organism is able to discriminate between self and nonself structures. Soybean (Glycine max L.) exploits a specific molecular pattern, a 1,6-␤-linked and 1,3-␤-branched heptaglucoside (HG), present in cell walls of the oomycetal pathogen Phytophthora sojae, as a signal compound eliciting the onset of defense reactions. The specific and high affinity HG-binding site is contained in the ␤-glucan-binding protein (GBP), which in turn is part of a proposed receptor complex. The ability to perceive and respond to Phytophthora cell wall-derived ␤-glucan elicitors is exclusive to plants that belong to the Fabaceae. However, we propose that the presence of the GBP is essential, but not sufficient for ␤-glucan elicitor-dependent disease resistance because genes encoding GBP-related proteins can be retrieved from many plant species. Furthermore, we show that the GBP is composed of two different carbohydrateactive protein domains, one containing the ␤-glucanbinding site, and the other related to glucan endoglucosidases of fungal origin. The glucan hydrolase displays most likely an endo-specific mode of action, cleaving only 1,3-␤-D-glucosidic linkages of oligoglucosides consisting of at least four moieties. Thus, the intrinsic endo-1,3-␤-glucanase activity of the GBP is perfectly suited during initial contact with Phytophthora to release oligoglucoside fragments enriched in motifs that constitute ligands for the high affinity binding site present in the same protein. The concept of innate immunity in plants receives substantial support by this highly sophisticated system using ancient enzyme modules as an active part of the recognition mechanism.

show abstract

Section: Methodsmentioning

confidence: 99%

An Ancient Enzyme Domain Hidden in the Putative β-Glucan Elicitor Receptor of Soybean May Play an Active Part in the Perception of Pathogen-associated Molecular Patterns during Broad Host Resistance

Fliegmann

Mithöfer

Wanner

et al. 2004

Journal of Biological Chemistry

148

115

View full text Add to dashboard Cite

show abstract

“…The Arabidopsis liquidgrown callus is a tissue that synthesizes a typical primary cell wall. Its cell wall is composed of cellulose, abundant pectic components, such as homogalacturonan and rhamnogalacturonan I, the hemicellulose xyloglucan, and the cell wall extensin proteins (Goubet et al, 2002;Handford et al, 2003;Manfield et al, 2004;Barton et al, 2006;P. Dupree, unpublished data).…”

Section: Quantities Of the Cell Wall Synthesis Machineries In The Golmentioning

confidence: 99%

Label-Free Protein Quantification for Plant Golgi Protein Localization and Abundance

et al. 2014

View full text Add to dashboard Cite

The proteomic composition of the Arabidopsis (Arabidopsis thaliana) Golgi apparatus is currently reasonably well documented; however, little is known about the relative abundances between different proteins within this compartment. Accurate quantitative information of Golgi resident proteins is of great importance: it facilitates a better understanding of the biochemical processes that take place within this organelle, especially those of different polysaccharide synthesis pathways. Golgi resident proteins are challenging to quantify because the abundance of this organelle is relatively low within the cell. In this study, an organelle fractionation approach targeting the Golgi apparatus was combined with a label-free quantitative mass spectrometry (dataindependent acquisition method using ion mobility separation known as LC-IMS-MS E [or HDMS E ]) to simultaneously localize proteins to the Golgi apparatus and assess their relative quantity. In total, 102 Golgi-localized proteins were quantified. These data show that organelle fractionation in conjunction with label-free quantitative mass spectrometry is a powerful and relatively simple tool to access protein organelle localization and their relative abundances. The findings presented open a unique view on the organization of the plant Golgi apparatus, leading toward unique hypotheses centered on the biochemical processes of this organelle.

show abstract

“…The protocol for the derivatization of sugar reducing ends using AMAC from Goubet et al [32] was applied to the oligo-λ-carrageenan series. Mixtures of DP (degree of polymerization) 4 and DP6 oligo-λ-carrageenans obtained after overnight enzymatic digestion (see above) were labelled according to the protocol described by Goubet et al [32].…”

Section: Synthesis and Enzymatic Digestion Of Fluorescent Oligosacchamentioning

confidence: 99%

“…Mixtures of DP (degree of polymerization) 4 and DP6 oligo-λ-carrageenans obtained after overnight enzymatic digestion (see above) were labelled according to the protocol described by Goubet et al [32]. After the grafting reaction had added the AMAC to the reducing ends, the samples were freezedried, dissolved in 50 mM (NH 4 ) 2 CO 3 , filtered and then purified by size-exclusion chromatography (Superdex 30 preparative grade column, 600 × 26 mm i.d.…”

Section: Synthesis and Enzymatic Digestion Of Fluorescent Oligosacchamentioning

confidence: 99%

“…The elution was performed in 50 mM (NH 4 ) 2 CO 3 running at 102 ml/h. The pure fluorescent oligosaccharides (DP4 and DP6) were collected, freeze-dried and stored at 4 • C. Fluorescent oligo-λ-carrabiose (DP2) was obtained using the protocol of Goubet et al [32] using purified DP2 as a substrate [6]. Since DP2 was eluted with salts and other small molecules, it could not be purified by sizeexclusion chromatography.…”

Section: Synthesis and Enzymatic Digestion Of Fluorescent Oligosacchamentioning

confidence: 99%

See 1 more Smart Citation

Degradation of λ-carrageenan by Pseudoalteromonas carrageenovora λ-carrageenase: a new family of glycoside hydrolases unrelated to κ- and ι-carrageenases

Guibet¹,

Colin²,

Barbeyron³

et al. 2007

Biochem. J.

View full text Add to dashboard Cite

Carrageenans are sulfated galactans found in the cell walls of red seaweeds. They are classified according to the number and the position of sulfate ester groups. lambda-Carrageenan is the most sulfated carrageenan and carries at least three sulfates per disaccharide unit. The sole known depolymerizing enzyme of lambda-carrageenan, the lambda-carrageenase from Pseudoalteromonas carrageenovora, has been purified, cloned and sequenced. Sequence analyses have revealed that the lambda-carrageenase, referred to as CglA, is the first member of a new family of GHs (glycoside hydrolases), which is unrelated to families GH16, that contains kappa-carrageenases, and GH82, that contains iota-carrageenases. This large enzyme (105 kDa) features a low-complexity region, suggesting the presence of a linker connecting at least two independent modules. The N-terminal region is predicted to fold as a beta-propeller. The main degradation products have been purified and characterized as neo-lambda-carratetraose [DP (degree of polymerization) 4] and neo-lambda-carrahexaose (DP6), indicating that CglA hydrolyses the beta-(1-->4) linkage of lambda-carrageenan. LC-MALLS (liquid chromatography-multi-angle laser light scattering) and (1)H-NMR monitoring of the enzymatic degradation of lambda-carrageenan indicate that CglA proceeds according to an endolytic mode of action and a mechanism of inversion of the anomeric configuration. Using 2-aminoacridone-labelled neo-lambda-carrabiose oligosaccharides, in the present study we demonstrate that the active site of CglA comprises at least 8 subsites (-4 to +4) and that a DP6 oligosaccharide binds in the subsites -4 to +2 and can be hydrolysed into DP4 and DP2.

show abstract

Polysaccharide Analysis Using Carbohydrate Gel Electrophoresis: A Method to Study Plant Cell Wall Polysaccharides and Polysaccharide Hydrolases

Cited by 167 publications

References 26 publications

An Ancient Enzyme Domain Hidden in the Putative β-Glucan Elicitor Receptor of Soybean May Play an Active Part in the Perception of Pathogen-associated Molecular Patterns during Broad Host Resistance

An Ancient Enzyme Domain Hidden in the Putative β-Glucan Elicitor Receptor of Soybean May Play an Active Part in the Perception of Pathogen-associated Molecular Patterns during Broad Host Resistance

Label-Free Protein Quantification for Plant Golgi Protein Localization and Abundance

Degradation of λ-carrageenan by Pseudoalteromonas carrageenovora λ-carrageenase: a new family of glycoside hydrolases unrelated to κ- and ι-carrageenases

Contact Info

Product

Resources

About