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Carbohydrate Gel Electrophoresis

  • Florence GoubetEmail author
  • Paul Dupree
  • Katja Salomon Johansen
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 715)

Abstract

Polysaccharide analysis using carbohydrate gel electrophoresis (PACE) relies on derivatization of the reducing ends of sugars with a fluorophore, followed by electrophoresis under optimized conditions in polyacrylamide gels. PACE is a sensitive and simple tool for studying polysaccharide structure or quantity and also has applications in the investigation of enzyme specificity.

Key words

Oligosaccharide Monosaccharide Polysaccharide Pectin Hemicellulose Enzyme 

References

  1. 1.
    Barton, C. J., Tailford, L., Welchman, H., Zhang, Z., Gilbert, H. J., Dupree, P., and Goubet, F. (2006) Enzymatic fingerprinting of Arabidopsis pectic polysaccharides using PACE-polysaccharide analysis by carbohydrate gel electrophoresis. Planta 224, 163–174.PubMedCrossRefGoogle Scholar
  2. 2.
    Handford, M. G., Baldwin, T. C., Goubet, F., Prime, T. A., Miles, J., Yu, X., and Dupree, P. (2003) Localisation and characterisation of mannan cell wall polysaccharides in Arabidopsis thaliana. Planta 218, 27–36.PubMedCrossRefGoogle Scholar
  3. 3.
    Brown, D. M., Goubet, F., Wong, V. W., Goodacre, R., Stephens, E., Dupree, P., and Turner, S. R. (2007) Comparison of five xylan synthesis mutants reveals new insight into the mechanisms of xylan synthesis. Plant J 52, 1154–1168.PubMedCrossRefGoogle Scholar
  4. 4.
    Karousou, E., Porta, G., De Luca, G., and Passi, A. (2004) Analysis of fluorophore-labelled hyaluronan and chondroitin sulfate disaccharides in biological samples. J Pharmac Bioch Anal 34, 791–795.CrossRefGoogle Scholar
  5. 5.
    Goubet, F., Morriswood, B., and Dupree, P. (2003) Analysis of methylated and unmethylated polygalacturonic acid structure by PACE: polysaccharide analysis using carbohydrate gel electrophoresis. Anal Biochem 321, 174–182.PubMedCrossRefGoogle Scholar
  6. 6.
    Goubet, F., Jackson, P., Deery, M., and Dupree, P. (2002) Polysaccharide analysis using carbohydrate gel electrophoresis (PACE): a method to study plant cell wall polysaccharides and polysaccharide hydrolases. Anal Biochem 300, 53–68.PubMedCrossRefGoogle Scholar
  7. 7.
    Fliegmann, J., Mithöfer, A., Wanner, G., and Ebel, J. (2004) 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. J Biol Chem 279, 1132–1140.PubMedCrossRefGoogle Scholar
  8. 8.
    Jackson, P. (1994) The analysis of fluorophore-labeled glycans by high-resolution polyacrylamide gel electrophoresis. Anal Biochem 216, 243–252.PubMedCrossRefGoogle Scholar
  9. 9.
    Goubet, F., Ström, A., Quéméner, B., Stephens, E., Williams, M. A. K., and Dupree, P. (2006) Resolution of the structural isomers of partially methylesterified oligogalacturonides by polysaccharide analysis using carbohydrate gel electrophoresis. Glycobiology 16, 29–35.PubMedCrossRefGoogle Scholar
  10. 10.
    Liners, F., Helbert, W., and Van Cutsem, P. (2005) Production and characterization of a phage-display recombinant antibody against carrageenans: evidence for the recognition of a secondary structure of carrageenan chains present in red algae tissues. Glycobiology 15, 849–860.PubMedCrossRefGoogle Scholar
  11. 11.
    Phalip, V., Goubet, F., Carapito, R., and Jeltsch, J. -M. (2009) Plant cell wall degradation with a powerful fusarium graminearum enzymatic arsenal. J Microbiol Biotechnol 19, 573–581.PubMedGoogle Scholar
  12. 12.
    Phalip, V., Delalande, F., Carapito, C., Goubet, F., Hatsch, D., Leize-Wagner, E., Dupree, P., Dorsselaer, A. V., and Jeltsch, J. M. (2005) Diversity of the exoproteome of Fusarium graminearum grown on plant cell wall. Curr Genet 48, 366–379.PubMedCrossRefGoogle Scholar
  13. 13.
    Palackal, N., Brennan, Y., Callen, N. C., Dupree, P., Frey, G., Goubet, F., Hazlewood, G. P., Healey, S., Kang, Y. E., Kretz, K. A., Lee, E., Tan, X., Tomlinson, G. L., Verruto, J., Wong, V. W. K., Mathur, E. J., Short, J. M., Robertson, D. E., and Steer, B. A. (2004) An evolutionary route to xylanase process fitness. Protein Sci 13, 494–503.PubMedCrossRefGoogle Scholar
  14. 14.
    Fanutti, C., Gidley, M. J., and Reid, G. J. S. (1996) Substrate subsite recognition of the xyloglucan endo-transglycosylase or xyloglucan-specific endo-(1→4)-β-d-glucanase from the cotyledons of germinated nasturtium (Tropaeolum majus L.) seeds. Planta 200, 221–228.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Florence Goubet
    • 1
    Email author
  • Paul Dupree
    • 2
  • Katja Salomon Johansen
    • 3
  1. 1.Bayer BioScience NVGhentBelgium
  2. 2.Department of BiochemistryUniversity of CambridgeCambridgeUK
  3. 3.Novozymes A/SBagsvaerdDenmark

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