Primary structure of the 2-O-methyl-α-l-fucose-containing side chain of the pectic polysaccharide, rhamnogalacturonan II

Glushka, John; Terrell, Mark; York, William S.; O’Neill, Malcolm A.; Gucwa, Angela L.; Darvill, Alan G.; Albersheim, Peter; Prestegard, James H.

doi:10.1016/s0008-6215(02)00461-5

Cited by 69 publications

(55 citation statements)

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“…By contrast, ARAf and XYL1, which release both D-Xyl and L-Ara, could be involved mainly in debranching the xylan and arabinoxylan polymers, by hydrolyzing the a-L-arabinofuranoside side groups. In addition, the results obtained suggest that in vivo ARAf and XYL1 are involved in the hydrolysis of arabinan side chains of RG-I, a polysaccharide of the primary cell wall (Ridley et al, 2001;Willats et al, 2001;Glushka et al, 2003). Arabidopsis antisense lines for XYL1 were reported to show alterations in plant development leading to a marked phenotype at the level of the leaf and silique shapes and a reduced height when compared to wildtype plants grown under the same growth conditions (Goujon et al, 2003).…”

Section: Discussionmentioning

confidence: 96%

“…Removal of side chain substituents in xylan is considered to be a limiting step in achieving efficient hydrolysis of the polysaccharide polymer (Rahman et al, 2003;Tuncer and Ball, 2003), since these groups create steric obstacles for the enzyme-substrate complex formation Rodionova, 1977a, 1977b). L-Ara, as a side chain substituent of xylan, is present in various plant structural polysaccharides such as rhamnogalacturonan-I (RG-I) and rhamnogalacturonan-II (Ridley et al, 2001;Willats et al, 2001;Glushka et al, 2003).…”

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Purification and Characterization of Enzymes Exhibiting β-d-Xylosidase Activities in Stem Tissues of Arabidopsis

et al. 2004

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This work describes the purification and characterization of enzymes that exhibit b-D-xylosidase activity in stem tissues of Arabidopsis. This is the first detailed investigation that concerns the characterization of catalytic properties and sequence identity of enzymes with b-D-xylosidase activities in a dicotyledonous plant. Three different enzymes, ARAf, XYL4, and XYL1 with apparent molecular masses of 75, 67, and 64 kD, respectively, were purified to homogeneity. ARAf was identified as a putative a-L-arabinofuranosidase, and XYL4 and XYL1 as putative b-D-xylosidases using matrix-assisted laser-desorption ionization time of flight. ARAf belongs to family 51 and XYL4 and XYL1 to family 3 of glycoside hydrolases. ARAf and XYL1 have highest specificity for p-nitrophenyl-a-L-arabinofuranoside and XYL4 for p-nitrophenyl-b-D-xylopyranoside and natural substrates such as xylobiose and xylotetraose. XYL4 was shown to release mainly D-Xyl from oat spelt xylan, rye arabinoxylan, wheat arabinoxylan, and oligoarabinoxylans. ARAf and XYL1 can also release D-Xyl from these substrates but less efficiently than XYL4. Moreover, they can also release L-Ara from arabinoxylans and arabinan. Overall, the results indicate that XYL4 possesses enzymatic specificity characteristic for a b-D-xylosidase, while ARAf and XYL1 act as bifunctional a-L-arabinofuranosidase/b-D-xylosidases. Analysis of the activity of these three enzymes in stem tissues at different stages of development has shown that young stems possess the highest activities for all three enzymes in comparison to the activities of the enzymes present in stems at older stages of development. High enzyme activities are most likely related to the necessary modifications of cell wall structure occurring during plant growth. Plant cells are surrounded by an extracellular matrix known as the cell wall. The regulation of cell wall morphology and composition is important for the determination of cell size and shape, cellular interactions with the environment, mechanical resistance, and defense against pathogen attacks (Reiter, 2002). The cell wall is also involved in plant growth and development (Stolle-Smits et al., 1999;Obel et al., 2002). In addition, although cell walls consist mainly of polysaccharides broadly classified as celluloses, hemicelluloses, and pectins, their constitution varies, not only from plant to plant, but also in different tissues of the same plant (Heredia et al., 1995;Cosgrove, 1997;Popper and Fry, 2003). A coordinated series of biochemical processes occur during plant development resulting in the biosynthesis and degradation of cell wall components. Consequently, numerous enzymes must be implicated in these processes (Heredia et al., 1995;Cosgrove, 1997;Stolle-Smits et al., 1999;Obel et al., 2002;Reiter, 2002;Popper and Fry, 2003). However, to date, little information is available concerning such enzymes, their mode of action, and their physiological role. Increased research interest has focused on enzymes involved in the metabolism of hemicelluloses in ...

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Section: Discussionmentioning

confidence: 96%

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confidence: 99%

Purification and Characterization of Enzymes Exhibiting β-d-Xylosidase Activities in Stem Tissues of Arabidopsis

et al. 2004

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“…In Arabidopsis cell walls, fucose is present in the hemicellulose xyloglucan (6), the pectins rhamnogalacturonan-I (RG-I) (25) and rhamnogalacturonan-II (RG-II) (26), arabinogalactan proteins (AGPs) (21), and N-linked glycoproteins (22). Each of these cell-wall components has a characteristic extraction pattern (27) and can be analytically digested by a specific set of enzymes (8).…”

Section: Biochemical Analysis Of Fucal Incorporation Into Arabidopsismentioning

confidence: 99%

Metabolic click-labeling with a fucose analog reveals pectin delivery, architecture, and dynamics in Arabidopsis cell walls

Anderson

Wallace

Somerville

2012

Proc. Natl. Acad. Sci. U.S.A.

137

133

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Polysaccharide-rich cell walls are a defining feature of plants that influence cell division and growth, but many details of cell-wall organization and dynamics are unknown because of a lack of suitable chemical probes. Metabolic labeling using sugar analogs compatible with click chemistry has the potential to provide new insights into cell-wall structure and dynamics. Using this approach, we found that an alkynylated fucose analog (FucAl) is metabolically incorporated into the cell walls of Arabidopsis thaliana roots and that a significant fraction of the incorporated FucAl is present in pectic rhamnogalacturonan-I (RG-I). Time-course experiments revealed that FucAl-containing RG-I first localizes in cell walls as uniformly distributed punctae that likely mark the sites of vesiclemediated delivery of new polysaccharides to growing cell walls. In addition, we found that the pattern of incorporated FucAl differs markedly along the developmental gradient of the root. Using pulse-chase experiments, we also discovered that the pectin network is reoriented in elongating root epidermal cells. These results reveal previously undescribed details of polysaccharide delivery, organization, and dynamics in cell walls.fluorescence | secretion | small molecule T he primary cell walls of plants consist of a complex polysaccharide-rich network containing cellulose, hemicellulose, pectin, and structural proteins. Cellulose, which is the primary load-bearing component of the wall, is hydrogen-bonded to hemicelluloses, and this composite is embedded in a pectin matrix (1, 2). The cell wall must simultaneously maintain the structural integrity of the plant cell by resisting the osmotic pressure necessary for turgor-mediated cell growth, and remain sufficiently dynamic to expand along with the growing cell. These requirements are satisfied by the controlled synthesis, deposition, and alteration of cell-wall components. Cellulose is synthesized at the plasma membrane (3), whereas hemicelluloses and pectins are synthesized intracellularly and secreted into the wall via an incompletely characterized vesicle-mediated trafficking pathway (2).In contrast to proteins, for which genetically encoded fluorescent tags have provided many insights into localization and function, detailed characterization of cell-wall polysaccharide structure and dynamics in intact plants has proven difficult. Chemical (4, 5) and biochemical (6-8) techniques can provide primary structural information for extracted cell-wall polymers. Imaging approaches, including transmission electron microscopy (9, 10), Raman microspectroscopy (11), and studies using a growing array of lectins, carbohydrate-binding modules, and carbohydrate-specific antibodies (12) have provided information regarding the location or abundance of cell-wall polysaccharides in various cell types (13). However, these imaging techniques suffer from limitations, including long sample-preparation times, fixation artifacts, lack of temporal information, lack of polymer specificity (12), and masking of a...

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“…[9] In particular, we were interested in whether the presence of this disaccharide moiety would affect the ring conformation of the crowded a-l-Arap residue. [11] The oligosaccharides were substituted with an artificial aminopropyl spacer to facilitate conjugation to carrier proteins, as required for immunization and ELISA.…”

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confidence: 99%

A Highly Convergent Chemical Synthesis of Conformational Epitopes of Rhamnogalacturonan II

Rao

Boons

2007

Angew Chem Int Ed

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Recent estimates indicate that more than 2000 genes encode proteins involved in the biosynthesis and remodeling of the polysaccharide-rich primary cell wall of plants.[1] The functions of only a handful of these gene products are known. [2,3] Furthermore, there is data to support the hypothesis that carbohydrate epitopes change during plant development; however, detailed knowledge about these processes is lacking. It is critical that these deficiencies in knowledge be addressed urgently because advances in plant sciences and genetics will provide researchers with tools to develop the next generation of agroenergy and agromaterial crops, which will be higheryielding and tailored for modern biorefinery operations. [4] Monoclonal antibodies are attractive tools for monitoring changes in the composition and organization of the primary plant wall at the cellular and subcellular level.[5] To date, only a small number of such monoclonal antibodies have been generated for which the binding specificities have been characterized. Synthetic chemistry can provide well-defined plant-derived oligosaccharides, [6][7][8] which can be employed for the production of monoclonal antibodies. In particular, the highly complex pectins are prime targets for this purpose. For example, rhamnogalacturonan II (RGII) is a structurally highly complex pectic oligosaccharide of the primary cell wall of higher plants [9] and is composed of four structurally different oligosaccharides referred to as side chains A-D, which are attached to a linear a(1!4)-linked d-GalpA polysaccharide backbone. These polysaccharides are crosslinked covalently through borate diesters (1:2) between two apiofuranosyl residues of the A side chain. The resulting three-dimensional pectic network contributes to the mechanical properties of the primary cell wall and is required for normal plant growth and development. Indeed, changes in cell-wall properties that decrease the borate cross-linking of pectin lead to symptoms such as dwarfism.[10]

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Primary structure of the 2-O-methyl-α-l-fucose-containing side chain of the pectic polysaccharide, rhamnogalacturonan II

Cited by 69 publications

References 37 publications

Purification and Characterization of Enzymes Exhibiting β-d-Xylosidase Activities in Stem Tissues of Arabidopsis

Purification and Characterization of Enzymes Exhibiting β-d-Xylosidase Activities in Stem Tissues of Arabidopsis

Metabolic click-labeling with a fucose analog reveals pectin delivery, architecture, and dynamics in Arabidopsis cell walls

A Highly Convergent Chemical Synthesis of Conformational Epitopes of Rhamnogalacturonan II

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