Structure of a Hydroxyproline (Hyp)-Arabinogalactan Polysaccharide from Repetitive Ala-Hyp Expressed in Transgenic Nicotiana tabacum

Li, Tan; Qiu, Feng; Lamport, Derek T. A.; Kieliszewski, Marcia J.

doi:10.1074/jbc.m311864200

Cited by 149 publications

(160 citation statements)

References 55 publications

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“…4), we propose that the two enzymes transfer Fuc residues onto two different Ara residues on AGP molecules (enzyme specificity dictated by the fucosylation sites). Tan et al (32) have shown that the expression of a synthetic gene encoding an [Ala-Hyp] 51 peptide in tobacco BY2 cells produced and secreted a mixture of glycoproteins having Hyp residues substituted with arabinogalactan (AG) polysaccharides (DP13-26) and containing terminal Ara residues ␣(1,3)-or ␣(1,5)-linked. A simplified presentation of this AG polysaccharide structure is depicted in Fig.…”

Section: Discussionmentioning

confidence: 99%

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Functional Identification of Two Nonredundant Arabidopsis α(1,2)Fucosyltransferases Specific to Arabinogalactan Proteins

Williams

Bernard

et al. 2010

Journal of Biological Chemistry

119

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Virtually nothing is known about the mechanisms and enzymes responsible for the glycosylation of arabinogalactan proteins (AGPs). The glycosyltransferase 37 family contains plant-specific enzymes, which suggests involvement in plantspecific organs such as the cell wall. Our working hypothesis is that AtFUT4 and AtFUT6 genes encode ␣(1,2)fucosyltransferases (FUTs) for AGPs. Multiple lines of evidence support this hypothesis. First, overexpression of the two genes in tobacco BY2 cells, known to contain nonfucosylated AGPs, resulted in a staining of transgenic cells with eel lectin, which specifically binds to terminal ␣-linked fucose. Second, monosaccharide analysis by high pH anion exchange chromatography and electrospray ionization mass spectrometry indicated the presence of fucose in AGPs from transgenic cell lines but not in AGPs from wild type cells. Third, detergent extracts from microsomal membranes prepared from transgenic lines were able to fucosylate, in vitro, purified AGPs from BY2 wild type cells. Susceptibility of [ 14 C]fucosylated AGPs to ␣(1,2)fucosidase, and not to ␣(1,3/4)fucosidase, indicated that an ␣(1,2) linkage is formed. Furthermore, dearabinosylated AGPs were not substrate acceptors for these enzymes, indicating that arabinosyl residues represent the fucosylation sites on these molecules. Testing of several polysaccharides, oligosaccharides, and glycoproteins as potential substrate acceptors in the fucosyl transfer reactions indicated that the two enzymes are specific for AGPs but are not functionally redundant because they differentially fucosylate certain AGPs. AtFUT4 and AtFUT6 are the first enzymes to be characterized for AGP glycosylation and further our understanding of cell wall biosynthesis.

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Section: Discussionmentioning

confidence: 99%

“…BY2 cells were chosen as the expression system for two reasons: (i) tobacco BY2 and Arabidopsis suspension cells produce similar AGPs (32), and (ii) AGPs in both BY2 and Arabidopsis suspension cells lack Fuc (32), making them ideal substrates for AGP-FUTs.…”

Section: ␣(12)futs Specific To Arabinogalactan Proteinsmentioning

confidence: 99%

Functional Identification of Two Nonredundant Arabidopsis α(1,2)Fucosyltransferases Specific to Arabinogalactan Proteins

Williams

Bernard

et al. 2010

Journal of Biological Chemistry

119

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“…These spheroidal nanoparticles are concentrated during evaporation, and the highly structural flexibility of the protein backbone as well as the anchored bulky AG branches of the AGPs allow these macromolecules to be tightly packed (29), reaching an intimate connection between adjacent nanoparticles. Subsequent Ca 2+ -driven cross-linking among carboxyl groups of the uronic acid residues within the AGPs and the pectic acids in the extracellular space favors the cohesion of the adjacent AGP-rich nanoparticles, gives rise to the generation of an adhesive film (24,31,38), further aids in the curing progress of the exuded adhesive, and eventually realizes the adhesive function at the interface by restraining the relative movement of the adventitious roots and the corresponding substrates. Throughout these procedures, the AGP-rich ivy nanoparticles also possess the capacity to permeate irregularities present on the substrates owing to their rough surfaces in most cases, resulting in a strong mechanical interlocking at the interface and further ensuring an ideal adhesive action (26,39,40,(42)(43)(44)(45)(46).…”

Section: Resultsmentioning

confidence: 99%

“…3A) (24,28,30,32,38). It has been demonstrated that numerous AGP molecules possess negative surface charge that is thought to be associated with the presence of uronic acid substituents (i.e., GlcA and GalA) at the termini of type II AGs (24,29,30).…”

Section: Resultsmentioning

confidence: 99%

Nanospherical arabinogalactan proteins are a key component of the high-strength adhesive secreted by English ivy

Huang

Wang

et al. 2016

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

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Over 130 y have passed since Charles Darwin first discovered that the adventitious roots of English ivy (Hedera helix) exude a yellowish mucilage that promotes the capacity of this plant to climb vertical surfaces. Unfortunately, little progress has been made in elucidating the adhesion mechanisms underlying this high-strength adhesive. In the previous studies, spherical nanoparticles were observed in the viscous exudate. Here we show that these nanoparticles are predominantly composed of arabinogalactan proteins (AGPs), a superfamily of hydroxyproline-rich glycoproteins present in the extracellular spaces of plant cells. The spheroidal shape of the AGP-rich ivy nanoparticles results in a low viscosity of the ivy adhesive, and thus a favorable wetting behavior on the surface of substrates. Meanwhile, calciumdriven electrostatic interactions among carboxyl groups of the AGPs and the pectic acids give rise to the cross-linking of the exuded adhesive substances, favor subsequent curing (hardening) via formation of an adhesive film, and eventually promote the generation of mechanical interlocking between the adventitious roots of English ivy and the surface of substrates. Inspired by these molecular events, a reconstructed ivy-mimetic adhesive composite was developed by integrating purified AGP-rich ivy nanoparticles with pectic polysaccharides and calcium ions. Information gained from the subsequent tensile tests, in turn, substantiated the proposed adhesion mechanisms underlying the ivy-derived adhesive. Given that AGPs and pectic polysaccharides are also observed in bioadhesives exuded by other climbing plants, the adhesion mechanisms revealed by English ivy may forward the progress toward understanding the general principles underlying diverse botanic adhesives.ivy nanoparticle | ivy adhesive | arabinogalactan protein | adhesion mechanism | reconstructed adhesive A lthough there is a growing interest in exploring mechanisms regulating a variety of adhesive behaviors in the animal kingdom (1-6), the molecular basis allowing creeping plants, such as English ivy (Hedera helix), to generate sufficient adhesive force, aiding in clinging to vertical surfaces, is rarely discussed (Fig. 1A). Previous studies have emphasized mechanical strategies exploited by multiple climbing organs that evolve in plants (7-11). Nevertheless, the role of the glue-like viscous exudates that are observed on the majority of these organs and that cement the plants to the substrates has been less explored (10,12,13). Diverse polysaccharides and glycoproteins, comprising mucilaginous pectins, arabinogalactans, arabinogalactan proteins (AGPs), and many others, have been identified to be the predominant components in these adhesive substances (14-17); however, the molecular mechanisms underlying the high-strength adhesion remain elusive.By means of atomic force microscopy (AFM), bulk spherical organic nanoparticles have been observed in the exudates derived from the root hairs of English ivy (18-20). These proteinaceous nanoparticles are presum...

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“…[1][2][3][4]35) In the present study, we used exo-β-(1→3)-galactanase to analyze specifically the distribution of L-Ara residues along the β-(1→6)-galactan side chains and their acidic derivatives capped with 4-MeGlcA. This yielded a variety of L-arabino-galactooligosaccharides, which are usually difficult to prepare by chemical degradation due to the acid-sensitive nature of their L-arabinofuranosidic linkages.…”

Section: Discussionmentioning

confidence: 99%

Enzymatic fragmentation of carbohydrate moieties of radish arabinogalactan-protein and elucidation of the structures

Shimoda

Okabe

Kotake

et al. 2014

Bioscience, Biotechnology, and Biochemistry

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We investigated the structures of L-arabino-galactooligosaccharides released from the sugar moieties of a radish arabinogalactan-protein (AGP) by the action of exo-β-(1→3)-galactanase. We detected a series of neutral β-(1→6)-linked galactooligosaccharides forming branches of one to up to at least 19 consecutive Gal groups, together with corresponding acidic derivatives terminating in 4-O-methyl-glucuronic acid (4-Me-GlcA) at the non-reducing end. Some oligosaccharide chains of degree of polymerization (dp) higher than 3 for neutral, and 4 for acidic oligomers were modified with L-Araf residues. The acidic tetrasaccharide 4-Me-β-GlcA-(1→6)[α-L-Araf-(1→3)]-β-Gal-(1→6)-Gal was detected as an abundant L-Araf-containing oligosaccharide among these neutral and acidic oligomers. A pentasaccharide containing an additional L-Araf group attached to the L-Ara in the tetrasaccharide through an α-(1→5)-linkage was also found. We observed L-arabino-galactooligosaccharides substituted with single or disaccharide L-Araf units at different Gal residues along these neutral and acidic β-(1→6)-galactooligosaccharide chains, indicating that these side chains are highly variable in length and substituted variously with L-Araf residues.

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Structure of a Hydroxyproline (Hyp)-Arabinogalactan Polysaccharide from Repetitive Ala-Hyp Expressed in Transgenic Nicotiana tabacum

Cited by 149 publications

References 55 publications

Functional Identification of Two Nonredundant Arabidopsis α(1,2)Fucosyltransferases Specific to Arabinogalactan Proteins

Functional Identification of Two Nonredundant Arabidopsis α(1,2)Fucosyltransferases Specific to Arabinogalactan Proteins

Nanospherical arabinogalactan proteins are a key component of the high-strength adhesive secreted by English ivy

Enzymatic fragmentation of carbohydrate moieties of radish arabinogalactan-protein and elucidation of the structures

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