Abstract
In eukaryotic cells, secreted proteins can be subjected to numerous post-translational modifications. One of these, glycosylation, consists of the addition of sugar residues to the protein backbone, while the protein enters or travels within the secretory pathway. Glycosylation of secreted proteins can be of two types∶ N- or O-glycosylation, depending on the linkage involved between the oligosaccharide moiety and the protein backbone.
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Abeijon, A. and Hirschberg, C. B. (1992) Topography of glycosylation reactions in the endoplasmic reticulum. Trends Biochem. Sci. 17, 32–36.
Szumilo, T., Kaushal, G. P., and Elbein, A. D. (1986) Purification and properties of a glucosidase I from mung bean seedlings. Arch. Biochem. Biophys. 247, 261–271.
Sturm, A., Johnson, K. D., Szumilo, T., Elbein, A. D., and Chrispeels, M. J. (1987) Subcellular localization of glycosidases and glycosyltransferases involved in the processing of N-linked oligosaccharides. Plant Physiol. 85, 741–745.
Trombetta, S. E., Bosch, M., and Parodi, A. J. (1991) Glucosylation of glycoproteins by mammalian, plant, fungal and trypanosomatid protozoa microsomal membranes. Biochemistry 28, 8108–8116.
Hammond, C., Braakman, I., and Helenius, A. (1994) Role of N-linked oligosaccharide recognition, glucose trimming, and calnexin in glycoprotein folding and quality control. Proc. Natl. Acad. Sci. USA 91, 913–917.
Huang, L., Franklin, A. E., and Hoffman, N. E. (1993) Primary structure and characterization of an Arabidopsis thaliana calnexin-like protein. J. Biol. Chem. 268, 6560–6566.
Lainé, A.-C., Gomord, V., and Faye, L. (1991) Xylose-specific antibodies as markers of subcompartmentation of terminal glycosylation in the Golgi apparatus of sycamore cells. FEBS Lett. 295, 179–184.
Zhang, G. F. and Staehelin, L. A. (1992) Functional compartmentation of the Golgi apparatus of plant cells. Plant Physiol. 99, 1070–1083.
Fitchette-Lainé, A..-C., Gomord, V., Chekkafi, A., and Faye, L. (1994) Distribution of xylosylation and fucosylation in the plant Golgi apparatus. Plant J. 5, 673–682.
Vitale, A. and Chrispeels, M. J. (1984) Transient N-acetylglucosamine in the biosynthesis of phytohemagglutinin∶ attachment in the Golgi apparatus and removal in the protein bodies. J. Cell Biol. 99, 133–140.
Takahashi, N., Hotta, T., Ishihara, H., Mori, M., Tejima, S., Bligny, R., Akazawa, T., Endo, S., and Arata, Y. (1986) Xylose-containing common structural unit in N-linked oligosaccharides of laccase from sycamore cells. Biochemistry 25, 388–395.
Schowalter, A. M. (1993) Structure and function of plant cell wall proteins. Plant Cell 5, 9–23.
Moore, P. J., Swords, K. M. M., Lynch, M. A., and Staehelin, L. A. (1991) Spatial organization of the assembly pathways of glycoproteins and complex polysaccha-rides in the Golgi apparatus of plants. J. Cell Biol. 112, 467–479.
Edge, A. S. B., Faltyneck, C. R., Hof, L., Le Reichet, J. R., and Weber, P. (1981). Deglycosylation of glycoproteins by trifluoromethane sulfonic acid. Anal. Biochem. 118, 131–137.
Rayon, C., Gomord, V., Faye, L., and Lerouge, P. (1996) N-glycosylation of bean and recombinant phytohemagglutinin. Plant Physiol. Biochem. 34, 273–281.
Shibuya, N., Goldstein, I. J., van Damme, E. J. M., and Peumans, W. J. (1988) Binding properties of a mannose-specific lectin from the snowdrop (Galanthus nivealis) bulb. J. Biol. Chem. 263, 728–734.
Animashaun, T., Mahmood, N., Hay, A. J., and Hughes, R. C. (1993) Inhibitory effect of novel mannose-binding lectins on HIV-infectivity and syncitium formation. Antiv. Chem. Chemother. 4, 145–153.
Faye, L. and Chrispeels, M. J. (1985) Characterization of N-linked oligosaccharides by affinoblotting with concanavalin A-peroxidase and treatment of the blots with glycosidases. Anal. Biochem. 149, 218–224.
Hsieh, P., Rosner, M. R., and Robbins, P. W. (1983) Selective clivage by endo β-N-acetylglucosaminidase H at individual glycosylation sites of Sindbis virion envelope glycoproteins. J. Biol. Chem. 258, 2555–2561.
Faye, L., Gomord, V., Fitchette-Lainé, A.-C., and Chrispeels, M. J. (1993) Affinity purification of antibodies specific for Asn-linked glycans containing α→3 fucose or β1→2 xylose. Anal. Biochem. 209, 104–108.
Lainé, A.-C. and Faye, L. (1988) Significant immunological cross-reactivity of plant glycoproteins. Electrophoresis 9, 841–844.
van Kuik, J. A., van Halbeek, H., Kamerling, J. P., and Vliegenthart, J. F. G. (1985) Primary structure of the low-molecular-weight carbohydrate chains of Helix pomatia α-hemocyanin. J. Biol. Chem. 260, 13,984–13,988.
Kubelka, V., Altmann, F., Staudacher, E., Tretter, V., März, L., Hard, K., Kamerling, J. P., and Vliegenthart, J. F. G. (1993) Primary structures of the N-linked carbohydrate chains from honeybee venom phospholipase A2. Eur. J. Biochem. 213, 1193–1204.
Kobata, A. (1979) Use of endo-and exoglycosidases for structural studies of glycoconjugates. Anal. Biochem. 100, 1–14.
Claradas, M. H. and Kaplan, A. (1984) Maturation of α-mannosidase in Dictyostelium discoideum∶ acquisition of endoglycosidase H resistance and sulfate. J. Biol. Chem. 259, 14,165–14,169.
Tretter, V., Altman, F., and März, L. (1991) Peptide-N4-(N-acetyl-β-glucosaminyl) asparagine amidase F cannot release glycans with fucose attached α 1–3 to the asparagine-linked N-acetylglucosamine residue. Eur. J. Biochem. 141, 97–104.
Woodward, J. R., Basic, A., Jahnen, W., and Clarke, A. E. (1989) N-linked glycan chains on S-allele-associated glycoproteins from Nicotiana alata. Plant Cell 1, 511–514.
D’Andrea, G., Salucci, M. L., Pitari, G., and Avigliano, L. (1993) Exhaustive removal of N-glycans from ascorbate oxidase∶ effect on the enzymatic activity and immunoreactivity. Glycobiology 3, 563–565.
Hori, H. and Elbein, A. D. (1981) Tunicamycin inhibits protein glycosylation in suspension cultured soybean cells. Plant Physiol. 67, 882–886.
Elbein, A. D. (1987) Inhibitors of the biosynthesis and processing of N-linked oligosaccharide chains. Annu. Rev. Biochem. 56, 497–537.
Driouich, A., Gonnet, P., Makkie, M., Lainé, A.-C., and Faye, L. (1989) The role of high-mannose and complex asparagine-linked glycans in the secretion and stability of glycoproteins. Planta 180, 96–104.
Bird, C. R., Gearing, A. J. H., and Thorpe, R. (1988) The use of Tween-20 alone as a blocking agent for the immunoblotting can cause artefactual results. J. Immunol. Meth. 106, 175–179.
Johnson, K. D. and Chrispeels, M. J. (1987) Substrate specificities of N-acetylglucosaminyl-, fucosyl-, and xylosyltransferases that modify glycoproteins in the Golgi apparatus of bean cotyledons. Plant Physiol. 84, 1301–1308.
Tezuka, K., Hayashi, M., Ishihara, H., Akazawa, T., and Takahashi, N. (1992) Studies on synthetic pathway of xylose-containing N-linked oligosaccharides deduced from substrate specificities of the processing enzymes in sycamore cells (Acer pseudoplatanus L.). Eur. J. Biochem. 203, 401–413.
Weitzhandler, M., KadWeitzhandler, M., Kadlecek, D., Avdalovic, N., Forte, J. G., Chow, D., and Townsend, R. R. (1993) Monosaccharide and oligosaccharide analysis of proteins transferred to polyvinylidene fluoride membranes after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. J. Biol. Chem. 268, 5121–5130.
Reiter, W.-D., Chapple, C. C. S., and Sommerville, C. R. (1993) Altered growth and cell walls in a fucose-deficient mutant of Arabidopsis. Science 261, 1032–1035.
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© 1998 Humana Press Inc., Totowa, NJ
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Fitchette-Lainé, AC., Denmat, LA., Lerouge, P., Faye, L. (1998). Analysis of N- and O-Glycosylation of Plant Proteins. In: Cunningham, C., Porter, A.J.R. (eds) Recombinant Proteins from Plants. Methods in Biotechnology, vol 3. Humana Press. https://doi.org/10.1007/978-1-60327-260-5_19
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DOI: https://doi.org/10.1007/978-1-60327-260-5_19
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Print ISBN: 978-0-89603-390-0
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