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Detergents and Chaotropes for Protein Solubilization before Two-Dimensional Electrophoresis

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Plant Proteomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 355))

Abstract

Because of the outstanding separating capabilities of two-dimensional electrophoresis for complete proteins, it would be advantageous to be able to apply it to all types of proteins. Unfortunately, severe solubility problems hamper the analysis of many classes of proteins, but especially membrane proteins. These problems arise mainly in the extraction and isoelectric focusing steps, and solutions are sought to improve protein solubility under the conditions prevailing during isoelectric focusing. These solutions deal mainly with chaotropes and new detergents, which are both able to enhance protein solubility. The input of these compounds in proteomics analysis of membrane proteins is discussed, as well as future directions.

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References

  1. Weber, K. and Kuter, D. J. (1971) Reversible denaturation of enzymes by sodium dodecyl sulfate. J. Biol. Chem. 246, 4504–4509.

    CAS  PubMed  Google Scholar 

  2. Ames, G. F. L. and Nikaido, K. (1976) Two-dimensional gel electrophoresis of membrane proteins. Biochemistry 15, 616–623.

    Article  CAS  PubMed  Google Scholar 

  3. Remy, R. and Ambard-Bretteville, F. (1987) Two dimensional electrophoresis in the analysis and preparation of cell organelle polypeptides. Methods Enzymol. 148, 623–632.

    Article  CAS  Google Scholar 

  4. Heizmann, C. W., Arnold, E. M., and Kuenzle, C. C. (1980) Fluctuations of non-histone chromosomal proteins in differentiating brain cortex and cerebellar neurons. J. Biol. Chem. 255, 11,504–11,511.

    CAS  PubMed  Google Scholar 

  5. Rabilloud, T., Adessi, C., Giraudel, A., and Lunardi, J. (1987) Improvement of the solubilization of proteins in two-dimensional electrophoresis with immobilized pH gradients. Electrophoresis 18, 307–316.

    Article  Google Scholar 

  6. Castellanos-Serra, L. and Paz-Lago, D. (2002) Inhibition of unwanted proteolysis during sample preparation: evaluation of its efficiency in challenge experiments. Electrophoresis 23, 1745–1753.

    Article  CAS  PubMed  Google Scholar 

  7. Santoni, V., Molloy, M., and Rabilloud, T. (2000) Membrane proteins and proteomics: un amour impossible? Electrophoresis 21, 1054–1070

    Article  CAS  PubMed  Google Scholar 

  8. Chevallet, M., Santoni, V., Poinas, A., et al. (1998) New zwitterionic detergents improve the analysis of membrane proteins by two-dimensional electrophoresis. Electrophoresis 19, 1901–1909.

    Article  CAS  PubMed  Google Scholar 

  9. Rabilloud, T., Blisnick, T., Heller, M., et al. (1999) Analysis of membrane proteins by two-dimensional electrophoresis: comparison of the proteins extracted from normal or Plasmodium falciparum-infected erythrocyte ghosts. Electrophoresis 20, 3603–3610

    Article  CAS  PubMed  Google Scholar 

  10. Tastet, C., Charmont, S., Chevallet, M., Luche, S., and Rabilloud, T. (2003) Structure-efficiency relationships of zwitterionic detergents as protein solubilizers in two-dimensional electrophoresis. Proteomics 3, 111–121.

    Article  CAS  PubMed  Google Scholar 

  11. Luche, S., Santoni, V., and Rabilloud, T. (2003) Evaluation of nonionic and zwitterionic detergents as membrane protein solubilizers in two-dimensional electrophoresis. Proteomics 3, 249–253.

    Article  CAS  PubMed  Google Scholar 

  12. Witzmann, F., Jarnot, B., and Parker, D. (1991) Dodecyl maltoside detergent improves resolution of hepatic membrane proteins in two-dimensional gels. Electrophoresis 12, 687–688.

    Article  CAS  PubMed  Google Scholar 

  13. Taylor, C. M. and Pfeiffer, S. E. (2003) Enhanced resolution of glycosylphosphatidylinositol-anchored and transmembrane proteins from the lipid-rich myelin membrane by two-dimensional gel electrophoresis. Proteomics 3, 1303–1312.

    Article  CAS  PubMed  Google Scholar 

  14. Santoni, V., Kieffer, S., Desclaux, D., Masson, F., and Rabilloud, T. (2000) Membrane proteomics: use of additive main effects with multiplicative interaction model to classify plasma membrane proteins according to their solubility and electrophoretic properties. Electrophoresis 21, 3329–3344.

    Article  CAS  PubMed  Google Scholar 

  15. Sanchez, J.C., Hochstrasser, D., and Rabilloud, T. (1999) In-gel sample rehydration of immobilized pH gradient. Methods Mol. Biol. 112, 221–225.

    CAS  PubMed  Google Scholar 

  16. MacFarlane, D.E. (1989) Two dimensional benzyldimethyl-n-hexadecylammonium chloride-sodium dodecyl sulfate preparative polyacrylamide gel electrophoresis: a high capacity high resolution technique for the purification of proteins from complex mixtures. Anal. Biochem. 176, 457–463.

    Article  CAS  PubMed  Google Scholar 

  17. Hartinger, J., Stenius, K., Hogemann, D., and Jahn, R. (1996) 16-BAC/SDS-PAGE: a two-dimensional gel electrophoresis system suitable for the separation of integral membrane proteins. Anal. Biochem. 240, 126–133.

    Article  CAS  PubMed  Google Scholar 

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© 2007 Humana Press Inc., Totowa, NJ

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Rabilloud, T., Luche, S., Santoni, V., Chevallet, M. (2007). Detergents and Chaotropes for Protein Solubilization before Two-Dimensional Electrophoresis. In: Thiellement, H., Zivy, M., Damerval, C., Méchin, V. (eds) Plant Proteomics. Methods in Molecular Biology, vol 355. Humana Press. https://doi.org/10.1385/1-59745-227-0:111

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  • DOI: https://doi.org/10.1385/1-59745-227-0:111

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-635-1

  • Online ISBN: 978-1-59745-227-4

  • eBook Packages: Springer Protocols

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