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Research ArticleBIOCHEMISTRY AND MACROMOLECULAR STRUCTURE
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Characterization of N-Glycans from Arabidopsis. Application to a Fucose-Deficient Mutant

Catherine Rayon, Marion Cabanes-Macheteau, Corinne Loutelier-Bourhis, Isabelle Salliot-Maire, Jérome Lemoine, Wolf-Dieter Reiter, Patrice Lerouge, Loı̈c Faye
Catherine Rayon
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Marion Cabanes-Macheteau
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Corinne Loutelier-Bourhis
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Isabelle Salliot-Maire
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Jérome Lemoine
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Wolf-Dieter Reiter
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Patrice Lerouge
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Loı̈c Faye
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Published February 1999. DOI: https://doi.org/10.1104/pp.119.2.725

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    Fig. 1.

    SDS-PAGE and western-blot analysis of protein extracts from leaves of wild-type Arabidopsis (lanes 1) ormur1 mutant (lanes 2). A, Silver staining; B, affinodetection with concanavalin A; C, immunodetection with antibodies specific for β(1,2)-Xyl-containing plant N-glycans; D, immunodetection with antibodies specific for α(1,3)-Fuc-containing plant N-glycans.

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    Fig. 2.

    HPAEC-PAD profile of high-mannose-typeN-glycans Man5GlcNAc2 to Man9GlcNAc2 (Man-5 to Man-9) isolated from wild-type Arabidopsis leaves. Corresponding structures are represented in Figure 3.

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    Fig. 3.

    Abbreviations and structures of high-mannose-typeN-glycans Man5GlcNAc2 to Man9GlcNAc2 (Man-5 to Man-9) isolated from wild-type Arabidopsis leaves.

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    Fig. 4.

    HPAEC profiles of the PA derivatives of paucimannosidic and complex-type N-glycans.N-glycans isolated from wild-type Arabidopsis (A) andmur1 mutant (B) leaves. The chromatograms presented in the boxed insets correspond to the C18 reverse-phase profiles of peaks b+c and f+g purified by HPAEC. Abbreviations refer to N-linked glycans represented in Figure 5.

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    Fig. 5.

    Abbreviations and structures of paucimannosidic and complex-type N-glycans isolated from Arabidopsis.

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    Fig. 6.

    HPAEC profiles of the β-N-acetylglucosaminidase digest of compound d after different incubation times. Peak a, PA derivative of M3XFGN2; peak b+c, PA derivatives of GN6-M3XFGN2 and GN3-M3XFGN2 (see Fig. 5).

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    Fig. 7.

    MS analysis of the mur1 N-glycans. A, Electrospray mass spectrum of the PA derivatives of the paucimannosidic and complex-type N-glycans isolated from the mur1 mutant. □, (M+H)+ ions of the PA derivatives of M3XGN2, GNM3XGN2, and GN2M3XGN2 (see Fig. 5). ♦, (M+H)+ ions of the PA derivatives of thel-Gal-containing oligosaccharides M3XGalGN2, GNM3XGalGN2, and GN2M3XGalGN2 (see Fig. 8). B, CID-MS-MS of the (M+H)+ ion of the PA derivative of M3XGalGN2 (m/z = 1283).

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    Fig. 8.

    Abbreviations and structures ofl-Gal-containing pauci-mannosidic and complex-typeN-glycans isolated from the mur1mutant.

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    Fig. 9.

    1H-NMR spectrum of the mixture of PA derivatives of the paucimanosidic and complex-typeN-glycans isolated from the mur1 mutant leaves. Numbers and letters refer to the corresponding protons (for example, 14 = signal of H-1 proton of residue 4). A, H1 signal of the nonsubstituted Man 4 of M3XGN2and GN6-M3XGN2. b, H1 signal of substituted Man 4 of GN3-M3XGN2 and GN2M3XGN2.  

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Characterization of N-Glycans from Arabidopsis. Application to a Fucose-Deficient Mutant
Catherine Rayon, Marion Cabanes-Macheteau, Corinne Loutelier-Bourhis, Isabelle Salliot-Maire, Jérome Lemoine, Wolf-Dieter Reiter, Patrice Lerouge, Loı̈c Faye
Plant Physiology Feb 1999, 119 (2) 725-734; DOI: 10.1104/pp.119.2.725

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Characterization of N-Glycans from Arabidopsis. Application to a Fucose-Deficient Mutant
Catherine Rayon, Marion Cabanes-Macheteau, Corinne Loutelier-Bourhis, Isabelle Salliot-Maire, Jérome Lemoine, Wolf-Dieter Reiter, Patrice Lerouge, Loı̈c Faye
Plant Physiology Feb 1999, 119 (2) 725-734; DOI: 10.1104/pp.119.2.725
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Plant Physiology: 119 (2)
Plant Physiology
Vol. 119, Issue 2
Feb 1999
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