SUMO Fusion Technology for Enhanced Protein Expression and Purification in Prokaryotes and Eukaryotes

  • Raymond J. Peroutka III
  • Steven J. Orcutt
  • James E. Strickler
  • Tauseef R. ButtEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 705)


The preparation of sufficient amounts of high-quality protein samples is the major bottleneck for structural proteomics. The use of recombinant proteins has increased significantly during the past decades. The most commonly used host, Escherichia coli, presents many challenges including protein misfolding, protein degradation, and low solubility. A novel SUMO fusion technology appears to enhance protein expression and solubility ( Efficient removal of the SUMO tag by SUMO protease in vitro facilitates the generation of target protein with a native N-terminus. In addition to its physiological relevance in eukaryotes, SUMO can be used as a powerful biotechnology tool for enhanced functional protein expression in prokaryotes and eukaryotes.

Key words

SUMO Smt3 SUMO protease1 protein expression protein solubility protein purification SUMOstar SUMOstar protease 


  1. 1.
    Seeler, J. S., Bischof, O., Nacerddine, K., Dejean, A. (2007) SUMO, the three Rs and cancer. Curr Top Microbiol Immunol 313, 49–71.PubMedCrossRefGoogle Scholar
  2. 2.
    Meinecke, I., Cinski, A., Baier, A., Peters, M. A., Dankbar, B., Wille, A., Drynda, A., Mendoza, H., Gay, R. E., Hay, R. T., Ink, B., Gay, S., Pap, T. (2007) Modification of nuclear PML protein by SUMO-1 regulates Fas-induced apoptosis in rheumatoid arthritis synovial fibroblasts. Proc Natl Acad Sci USA 104, 5073–5078.PubMedCrossRefGoogle Scholar
  3. 3.
    Rajan, S., Plant, L. D., Rabin, M. L., Butler, M. H., Goldstein, S. A. (2005) Sumoylation silences the plasma membrane leak K+ channel K2P1. Cell 121, 37–47.PubMedCrossRefGoogle Scholar
  4. 4.
    Martin, S., Nishimune, A., Mellor, J. R., Henley, J. M. (2007) SUMOylation regulates kainate-receptor-mediated synaptic transmission. Nature 447, 321–325.PubMedCrossRefGoogle Scholar
  5. 5.
    Mabb, A. M., Wuerzberger-Davis, S. M., Miyamoto, S. (2006) PIASy mediates NEMO sumoylation and NF-kappaB activation in response to genotoxic stress. Nat Cell Biol 8, 986–993.PubMedCrossRefGoogle Scholar
  6. 6.
    Li, S. J., Hochstrasser, M. (1999) A new protease required for cell-cycle progression in yeast. Nature 398, 246–251.PubMedCrossRefGoogle Scholar
  7. 7.
    Malakhov, M. P., Mattern, M. R., Malakhova, O. A., Drinker, M., Weeks, S. D., Butt, T. R. (2004) SUMO fusions and SUMO-specific protease for efficient expression and purification of proteins. J Struct Funct Genomics 5, 75–86.PubMedCrossRefGoogle Scholar
  8. 8.
    Marblestone, J. G., Edavettal, S. C., Lim, Y., Lim, P., Zuo, X., Butt, T. R. (2006) Comparison of SUMO fusion technology with traditional gene fusion systems: enhanced expression and solubility with SUMO. Protein Sci 15, 182–189.PubMedCrossRefGoogle Scholar
  9. 9.
    Butt, T. R., Edavettal, S. C., Hall, J. P., Mattern, M. R. (2005) SUMO fusion technology for difficult-to-express proteins. Protein Expr Purif 43, 1–9.PubMedCrossRefGoogle Scholar
  10. 10.
    Zuo, X., Li, S., Hall, J., Mattern, M. R., Tran, H., Shoo, J., Tan, R., Weiss, S. R., Butt, T. R. (2005) Enhanced expression and purification of membrane proteins by SUMO fusion in Escherichia coli. J Struct Funct Genomics 6, 103–11.PubMedCrossRefGoogle Scholar
  11. 11.
    Zuo, X., Mattern, M. R., Tan, R., Li, S., Hall, J., Sterner, D. E., Shoo, J., Tran, H., Lim, P., Sarafianos, S. G., Kazi, L., Navas-Martin, S., Weiss, S. R., Butt, T. R. (2005) Expression and purification of SARS coronavirus proteins using SUMO-fusions. Protein Expr Purif 42, 100–110.PubMedCrossRefGoogle Scholar
  12. 12.
    Guzzo, C. M., Yang, D. C. (2007) Systematic analysis of fusion and affinity tags using human aspartyl-tRNA synthetase expressed in E. coli. Protein Expr Purif 54, 166–175.PubMedCrossRefGoogle Scholar
  13. 13.
    Dominy, J. E., Jr., Simmons, C. R., Hirschberger, L. L., Hwang, J., Coloso, R. M., Stipanuk, M. H. (2007) Discovery and characterization of a second mammalian thiol dioxygenase, cysteamine dioxygenase. J Biol Chem 282, 25189–25198.PubMedCrossRefGoogle Scholar
  14. 14.
    Liu, L., Spurrier, J., Butt, T. R., Strickler, J. E. (2008) Enhanced protein expression in the baculovirus/insect cell system using engineered SUMO fusions. Protein Expr Purif, 62, 21–28.Google Scholar
  15. 15.
    Peroutka, R. J., Elshourbagy, N., Piech, T., Butt, T. R. (2008) Enhanced protein expression in mammalian cells using engineered SUMO fusions: secreted phospholipase A2. Protein Sci 17, 1586–1595.PubMedCrossRefGoogle Scholar
  16. 16.
    Mossessova, E., Lima, C. D. (2000) Ulp1-SUMO crystal structure and genetic analysis reveal conserved interactions and a regulatory element essential for cell growth in yeast. Mol Cell 5, 865–876.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Raymond J. Peroutka III
    • 1
  • Steven J. Orcutt
    • 1
  • James E. Strickler
    • 1
  • Tauseef R. Butt
    • 1
    Email author
  1. 1.LifeSensors Inc.MalvernUSA

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