Skip to main content

Advertisement

SpringerLink
Log in

Techniques for monitoring protein misfolding and aggregation in vitro and in living cells

  • Invited Review Paper
  • Published:
Korean Journal of Chemical Engineering Aims and scope Submit manuscript

Abstract

Protein misfolding and aggregation have been considered important in understanding many neurodegenerative diseases and recombinant biopharmaceutical production. Various traditional and modern techniques have been utilized to monitor protein aggregation in vitro and in living cells. Fibril formation, morphology and secondary structure content of amyloidogenic proteins in vitro have been monitored by molecular probes, TEM/AFM, and CD/FTIR analyses, respectively. Protein aggregation in living cells has been qualitatively or quantitatively monitored by numerous molecular folding reporters based on either fluorescent protein or enzyme. Aggregation of a target protein is directly correlated to the changes in fluorescence or enzyme activity of the folding reporter fused to the target protein, which allows non-invasive monitoring aggregation of the target protein in living cells. Advances in the techniques used to monitor protein aggregation in vitro and in living cells have greatly facilitated the understanding of the molecular mechanism of amyloidogenic protein aggregation associated with neurodegenerative diseases, optimizing culture conditions to reduce aggregation of biopharmaceuticals expressed in living cells, and screening of small molecule libraries in the search for protein aggregation inhibitors.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. M. Bartolini and V. Andrisano, ChemBioChem, 11, 1018 (2010).

    Article  CAS  Google Scholar 

  2. D.W. Colby, J. P. Cassady, G. C. Lin, V. M. Ingram and K.D. Wittrup, Nat. Chem. Biol., 2, 319 (2006).

    Article  CAS  Google Scholar 

  3. B. Keshet, J. J. Gray and T. A. Good, Protein Sci., 19, 2291 (2010).

    Article  CAS  Google Scholar 

  4. S. Lee, E. J. Fernandez and T. A. Good, Protein Sci., 16, 723 (2007).

    Article  CAS  Google Scholar 

  5. R. M. Murphy, Annu. Rev. Biomed. Eng., 4, 155 (2002).

    Article  CAS  Google Scholar 

  6. C. J. Pike, A. J. Walencewicz, C.G. Glabe and C.W. Cotman, Brain Research, 563, 311 (1991).

    Article  CAS  Google Scholar 

  7. A. Alzheimer’s, Alzheimer’s & Dementia, 6, 158 (2010).

    Article  Google Scholar 

  8. D. Majoor-Krakauer, P. J. Willems and A. Hofman, Clin. Genet., 63, 83 (2003).

    Article  CAS  Google Scholar 

  9. T. Kabuta, Y. Suzuki and K. Wada, J. Biol. Chem., 281, 30524 (2006).

    Article  CAS  Google Scholar 

  10. J. Niwa, S. Yamada, S. Ishigaki, J. Sone, M. Takahashi, M. Katsuno, F. Tanaka, M. Doyu and G. Sobue, J. Biol. Chem., 282, 28087 (2007).

    Article  CAS  Google Scholar 

  11. N. Ferrer-Miralles, J. Domingo-Espin, J. Corchero, E. Vazquez and A. Villaverde, Microbial Cell Factories, 8, 17 (2009).

    Article  CAS  Google Scholar 

  12. K. Graumann and A. Premstaller, Biotechnol. J., 1, 164 (2006).

    Article  CAS  Google Scholar 

  13. B. Jurgen, A. Breitenstein, V. Urlacher, K. Buttner, H, Lin, M. Hecker, T. Schweder and P. Neubauer, Microbial Cell Factories, 9, 41 (2010).

    Article  CAS  Google Scholar 

  14. J. G. Thomas and F. Baneyx, J. Biol. Chem., 271, 11141 (1996).

    Article  CAS  Google Scholar 

  15. Y. A. Ioannou, D. F. Bishop and R. J. Desnick, J. Cell Biol., 119, 1137 (1992).

    Article  CAS  Google Scholar 

  16. S. Ishii, R. Kase, T. Okumiya, H. Sakuraba and Y. Suzuki, Biochem. Biophys. Res. Commun., 220, 812 (1996).

    Article  CAS  Google Scholar 

  17. M. Schröder, R. Schäfer and P. Friedl, Biotechnol. Bioeng., 78, 131 (2002).

    Article  CAS  Google Scholar 

  18. T. Okumiya, M. A. Kroos, L. Van Wet, H. Takeuchi, A. T. Van der Ploeg and A. J. J. Reuser, Mol. Genet. Metab., 90, 49 (2007).

    Article  CAS  Google Scholar 

  19. S.-J. Hwang, S. Yoon, G. Koh and G. Lee, Appl. Microbiol. Biotechnol., 91, 305 (2011).

    Article  CAS  Google Scholar 

  20. N. Jenkins, P. Meleady, R. Tyther and L. Murphy, Biotechnol. Appl. Biochem., 53, 73 (2009).

    Article  CAS  Google Scholar 

  21. G. S. Waldo, B. M. Standish, J. Berendzen and T.C. Terwilliger, Nat. Biotechnol., 17, 691 (1999).

    Article  CAS  Google Scholar 

  22. J. A. H. G. Hardy, Science, 256, 184 (1992).

    Article  CAS  Google Scholar 

  23. R. Kayed, E. Head, J. L. Thompson, T.M. McIntire, S. C. Milton, C.W. Cotman and C.G. Glabe, Science (New York), 300, 486 (2003).

    Article  CAS  Google Scholar 

  24. C.G. Glabe, J. Biol. Chem., 283, 29639 (2008).

    Article  CAS  Google Scholar 

  25. M. Bucciantini, E. Giannoni, F. Chiti, F. Baroni, L. Formigli, J. Zurdo, N. Taddei, G. Ramponi, C.M. Dobson and M. Stefani, Nature, 416, 507 (2002).

    Article  CAS  Google Scholar 

  26. J. P. Cleary, D. M. Walsh, J. J. Hofmeister, G. M. Shankar, M. a. Kuskowski, D. J. Selkoe and K. H. Ashe, Nat. Neurosci., 8, 79 (2005).

    Article  CAS  Google Scholar 

  27. A. A. Reinke and J.E. Gestwicki, Chemical Biology & Drug Design, 70, 206 (2007).

    Article  CAS  Google Scholar 

  28. M. R. Nilsson, Methods, 34, 151 (2004).

    Article  CAS  Google Scholar 

  29. M. Groenning, J. Chem. Biol., 1 (2009).

  30. I. Maezawa, H.-S. Hong, R. Liu, C.-Y. Wu, R. H. Cheng, M.-P. Kung, H. F. Kung, K. S. Lam, S. Oddo, F.M. Laferla and L.-W. Jin, J. Neurochem., 104, 457 (2008).

    CAS  Google Scholar 

  31. S. A. Hudson, H. Ecroyd, T.W. Kee and J. A. Carver, The FEBS Journal, 276, 5960 (2009).

    Article  CAS  Google Scholar 

  32. A. A. Reinke and J.E. Gestwicki, Chemical Biology & Drug Design, 77, 399 (2011).

    Article  CAS  Google Scholar 

  33. M. Biancalana and S. Koide, Biochim. Biophys. Acta, 1804, 1405 (2010).

    CAS  Google Scholar 

  34. D. M. Hatters and M. D.W. Griffin, Amyloid International Journal of Experimental and Clinical Investigation, 752, 121 (2011).

    CAS  Google Scholar 

  35. M. Lindgren and P. Hammarström, The FEBS Journal, 277, 1380 (2010).

    Article  CAS  Google Scholar 

  36. H. E. Wong and I. Kwon, Plos One, 6, e25752 (2011).

    Article  CAS  Google Scholar 

  37. A. R. A. Ladiwala, J. S. Dordick and P.M. Tessier, J. Biol. Chem., 286, 3209 (2011).

    Article  CAS  Google Scholar 

  38. G. Bhak, J.-H. Lee, J.-S. Hahn and S.R. Paik, Plos One, 4, e4177 (2009).

    Article  CAS  Google Scholar 

  39. J.-H. Lee, G. Bhak, S.-G. Lee and S. R. Paik, Biophys. J., 95, L16 (2008).

    Article  CAS  Google Scholar 

  40. J. Ryu, M. Kanapathipillai, G. Lentzen and C. B. Park, Peptides, 29, 578 (2008).

    Article  CAS  Google Scholar 

  41. J.W. Wu, L. Breydo, J. M. Isas, J. Lee, Y.G. Kuznetsov, R. Langen and C. Glabe, J. Biol. Chem., 285, 6071 (2010).

    Article  CAS  Google Scholar 

  42. M. Ahmed, J. Davis, D. Aucoin, T. Sato, S. Ahuja, S. Aimoto, J. I. Elliott, W. E. Van Nostrand and S. O. Smith, Nat. Struct. Mol. Biol., 17, 561 (2010).

    Article  CAS  Google Scholar 

  43. A. R. A. Ladiwala, J. C. Lin, S. S. Bale, A. M. Marcelino-Cruz, M. Bhattacharya, J. S. Dordick and P.M. Tessier, J. Biol. Chem., 285, 24228 (2010).

    Article  CAS  Google Scholar 

  44. W. E. Klunk, R. F. Jacob and R. P. Mason, Methods Enzymol., 309, 6879 (1999).

    Google Scholar 

  45. D.M. Walsh, D.M. Hartley, Y. Kusumoto, Y. Fezoui, M.M. Condron, A. Lomakin, G. B. Benedek, D. J. Selkoe, D.B. Teplow and D. B. J. Biol, J. Biol. Chem., 274, 25945 (1999).

    Article  CAS  Google Scholar 

  46. A. Hawe, M. Sutter and W. Jiskoot, Pharmaceutical Research, 25, 1487 (2008).

    Article  CAS  Google Scholar 

  47. C. G. Glabe, The Open Biology Journal, 2, 222 (2009).

    Article  CAS  Google Scholar 

  48. A. R. A. Ladiwala, J.M. Perchiacca, Z. S. Fishman, M. Bhattacharya, A. M. Hickey, B.G. Domigan, J. S. Dordick, and P.M. Tessier, Biotechnology and Bioengineering, 109, 1869 (2012).

    Article  CAS  Google Scholar 

  49. S. Campioni, B. Mannini, M. Zampagni, A. Pensalfini, C. Parrini, E. Evangelisti, A. Relini, M. Stefani, C.M. Dobson, C. Cecchi and F. Chiti, Nat. Chem. Biol., 6, 140 (2010).

    Article  CAS  Google Scholar 

  50. B. Bolognesi, J. R. Kumita, T. P. Barros, E. K. Esbjorner, L.M. Luheshi, D. C. Crowther, M. R. Wilson, C. M. Dobson, G. Favrin and J. J. Yerbury, ACS Chem. Biol., 5, 735 (2010).

    Article  CAS  Google Scholar 

  51. R. Kayed, E. Head, F. Sarsoza, T. Saing, C.W. Cotman, M. Necula, L. Margol, J. Wu, L. Breydo, J. L. Thompson, S. Rasool, T. Gurlo, P. Butler and C. G. Glabe, Mol. Neurodegener., 2, 18 (2007).

    Article  CAS  Google Scholar 

  52. R. Kayed, A. Pensalfini, L. Margol, Y. Sokolov, F. Sarsoza, E. Head, J. Hall and C. Glabe J. Biol. Chem., 284, 4230 (2009).

    Article  CAS  Google Scholar 

  53. H. LeVine, III and L. C. Walker, Neurobiology of Aging, 31, 542 (2010).

    Article  CAS  Google Scholar 

  54. M. Fandrich, J. Mol. Biol., http://dx.doi.org/10.1016/j.jmb.2012.01.006 (2012).

  55. C. C. Glabe, Subcellular Biochemistry, 38, 167 (2005).

    Article  CAS  Google Scholar 

  56. H. E. Wong, W. Qi, H.-M. Choi, E. J. Fernandez and I. Kwon, ACS Chem. Neurosci., 2, 645 (2011).

    Article  CAS  Google Scholar 

  57. E. Cerf, R. Sarroukh, S. Tamamizu-Kato, L. Breydo, S. Derclaye, Y. F. Dufrêne, V. Narayanaswami, E. Goormaghtigh, J.-M. Ruysschaert and V. Raussens, Biochem. J., 421, 415 (2009).

    Article  CAS  Google Scholar 

  58. R. Sarroukh, E. Cerf, S. Derclaye, Y. F. Dufrêne, E. Goormaghtigh, J.-M. Ruysschaert and V. Raussens, Cellular and Molecular Life Sciences: CMLS, 68, 1429 (2011).

    Article  CAS  Google Scholar 

  59. Y. Feng, X.-P. Wang, S.-G. Yang, Y.-J. Wang, X. Zhang, X.-T. Du, X.-X. Sun, M. Zhao, L. Huang and R.-t. Liu, Neurotoxicology, 30, 986 (2009).

    Article  CAS  Google Scholar 

  60. L. A. Woods, G.W. Platt, A. L. Hellewell, E.W. Hewitt, S.W. Homans, A. E. Ashcroft and S. E. Radford, Nat. Chem. Biol., 7, 730 (2011).

    Article  CAS  Google Scholar 

  61. M. Necula, R. Kayed, S. Milton and C.G. Glabe, J. Biol. Chem., 282, 10311 (2007).

    Article  CAS  Google Scholar 

  62. X.-p. Wang, J.-h. Zhang, Y.-j. Wang, Y. Feng, X. Zhang, X.-x. Sun, J.-l Li, X.-t. Du, M. P. Lambert, S.-g. Yang, M. Zhao, W. L. Klein and R.-t. Liu, FEBS Lett., 583, 579 (2009).

    Article  CAS  Google Scholar 

  63. R. Kayed, I. Canto, L. Breydo, S. Rasool, T. Lukacsovich, J. Wu, R. Albay, A. Pensalfini, S. Yeung, E. Head, J. L. Marsh and C. Glabe, Mol. Neurodegener., 5, 57 (2010).

    Article  CAS  Google Scholar 

  64. A. Sood, M. Abid, C. Sauer, S. Hailemichael, M. Foster, B. Török and M. Török, Bioorg. Med. Chem. Lett., 21, 2044 (2011).

    Article  CAS  Google Scholar 

  65. D. Lee, E.-K. Lee, J.-H. Lee, C.-S. Chang and S. R. Paik, Eur. J. Biochem., 268, 295 (2001).

    Article  CAS  Google Scholar 

  66. C. Ha and C. B. Park, Biotechnol. Bioeng., 90, 848 (2005).

    Article  CAS  Google Scholar 

  67. S. H. Ku and C. B. Park, Langmuir, 24, 13822 (2008).

    Article  CAS  Google Scholar 

  68. R. Friedman, Biochem. J., 438, 415 (2011).

    CAS  Google Scholar 

  69. K.A. Dasilva, J. E. Shaw and J. McLaurin, Exp. Neurol., 223, 311 (2010).

    Article  CAS  Google Scholar 

  70. S. L. Gras, L. J. Waddington and K. N. Goldie, Notes, 752, 197 (2011).

    CAS  Google Scholar 

  71. B.H. Toyama and J. S. Weissman, Annu. Rev. Biochem., 80, 557 (2011).

    Article  CAS  Google Scholar 

  72. A. E. Langkilde and B. Vestergaard, FEBS Lett., 583, 2600 (2009).

    Article  CAS  Google Scholar 

  73. S. Chimon, M. A. Shaibat, C. R. Jones, D.C. Calero, B. Aizezi and Y. Ishii, Nat. Struct. Mol. Biol., 14, 1157 (2007).

    Article  CAS  Google Scholar 

  74. W. B. Stine, K. N. Dahlgren, G. a. Krafft and M. J. LaDu, The Journal of Biological Chemistry, 278, 11612 (2003).

    Article  CAS  Google Scholar 

  75. A. Perczel, K. Park and G. D. Fasman, Proteins: Structure, Function, and Bioinformatics, 13, 57 (1992).

    Article  CAS  Google Scholar 

  76. L. Whitmore and B. A. Wallace, Biopolymers, 89, 392 (2008).

    Article  CAS  Google Scholar 

  77. M. Bartolini, C. Bertucci, M. L. Bolognesi, A. Cavalli, C. Melchiorre and V. Andrisano, Chembiochem: A European Journal of Chemical Biology, 8, 2152 (2007).

    Article  CAS  Google Scholar 

  78. N. J. Greenfield, Trends in Analytical Chemistry, 18, 236 (1999).

    Article  CAS  Google Scholar 

  79. J. T. Pelton and L. R. McLean, Anal. Biochem., 277, 167 (2000).

    Article  CAS  Google Scholar 

  80. W. K. Surewicz, J. H. H. Mantsch and D. Chapman, Biochemistry, 32 (1993).

  81. D. H. A. Corrêa and C. H. I. Ramos, J. Biochem., 3, 164 (2009).

    Google Scholar 

  82. C. Soto and E. M. Castan, J. Biochem., 314, 701 (1996).

    CAS  Google Scholar 

  83. E. P. Vieira, H. Hermel and H. Möhwald, Biochim. Biophys. Acta, 1645, 6 (2003).

    CAS  Google Scholar 

  84. S. Tomaselli, V. Esposito, P. Vangone, N. a. J. van Nuland, A.M. J. J. Bonvin, R. Guerrini, T. Tancredi, P. a. Temussi and D. Picone, Chembiochem: A European Journal of Chemical Biology, 7, 257 (2006).

    Article  CAS  Google Scholar 

  85. N. J. Greenfield, Nature Protocols, 1, 2876 (2006).

    Article  CAS  Google Scholar 

  86. W. Kim and M. H. Hecht, J. Biol. Chem., 280, 35069 (2005).

    Article  CAS  Google Scholar 

  87. W. Kim and M. H. Hecht, J. Mol. Biol., 377, 565 (2008).

    Article  CAS  Google Scholar 

  88. P.E. Arslan and A. Chakrabartty, Biochem. Cell Biol., 87, 631 (2009).

    Article  CAS  Google Scholar 

  89. P. E. Arslan, V.K. Mulligan, S. Ho and A. Chakrabartty, J. Mol. Biol., 396, 1284 (2010).

    Article  CAS  Google Scholar 

  90. C. Heddle and S. L. Mazaleyrat, Protein. Eng. Des. Sel., 20, 327 (2007).

    Article  CAS  Google Scholar 

  91. S. Gregoire and I. Kwon, ACS Meeting, 241, 234–BIOT (2011).

    Google Scholar 

  92. W. Kim, Y. Kim, J. Min, D. J. Kim, Y. T. Chang and M.H. Hecht, ACS Chem. Biol., 1, 461 (2006).

    Article  CAS  Google Scholar 

  93. J. Geng, M. Li, J. Ren, E. Wang and X. Qu, Angew. Chem. Int. Ed. Eng., 50, 4184 (2011).

    Article  CAS  Google Scholar 

  94. B. Philipps, J. Hennecke and R. Glockshuber, J. Mol. Biol., 327, 239 (2003).

    Article  CAS  Google Scholar 

  95. S. Cabantous, T. C. Terwilliger and G. S. Waldo, Nat. Biotechnol., 23, 102 (2005).

    Article  CAS  Google Scholar 

  96. S. Cabantous, J. D. Pédelacq, B. L. Mark, C. Naranjo, T. C. Terwilliger and G. S. Waldo, J. Struct. Funct. Genomics., 6, 113 (2005).

    Article  CAS  Google Scholar 

  97. S. Cabantous and G. S. Waldo, Nat. Methods, 3, 845 (2006).

    Article  CAS  Google Scholar 

  98. W. J. Chun, G. S. Waldo and G.V.W. Johnson, J. Neurochem., 103, 2529 (2007).

    Article  CAS  Google Scholar 

  99. P. Listwan, T.C. Terwilliger and G. S. Waldo, J. Struct. Funct. Genomics., 10, 47 (2009).

    Article  CAS  Google Scholar 

  100. M.A. Lockard, P. Listwan, J.D. Pedelacq, S. Cabantous, H.B. Nguyen, T. C. Terwilliger and G. S. Waldo, Protein Engineering Design & Selection, 24, 565 (2011).

    Article  CAS  Google Scholar 

  101. K. L. Maxwell, A.K. Mittermaier, J. D. Forman-Kay and A. R. Davidson, Protein Sci., 8, 1908 (1999).

    Article  CAS  Google Scholar 

  102. M.R. Dyson, R.L. Perera, S. P. Shadbolt, L. Biderman, K. Bromek, N.V. Murzina and J. McCafferty, Nucleic Acids Res., 36, e51. Epub 2008 Apr 17. (2008).

    Article  CAS  Google Scholar 

  103. M. R. Dyson, Biochem. Soc. Trans., 38, 908 (2010).

    Article  CAS  Google Scholar 

  104. M. Morell, N. S. de Groot, J. Vendrell, F.X. Aviles and S. Ventura, Mol. Biosyst., 7, 1121 (2011).

    Article  CAS  Google Scholar 

  105. W.C. Wigley, R.D. Stidham, N.M. Smith, J. F. Hunt and P. J. Thomas, Nat. Biotechnol., 19, 131 (2001).

    Article  CAS  Google Scholar 

  106. L. Foit, G. J. Morgan, M. J. Kern, L.R. Steimer, A.A. von Hacht, J. Titchmarsh, S. L. Warriner, S. E. Radford and J.C. Bardwell, Mol. Cell., 36, 861 (2009).

    Article  CAS  Google Scholar 

  107. T. J. Mansell, S.W. Linderman, A.C. Fisher and M. P. DeLisa, Protein Science, 19, 1079 (2010).

    Article  CAS  Google Scholar 

  108. T. J. Mansell, A.C. Fisher and M. P. DeLisa, Curr. Protein Peptide Sci., 9, 138 (2008).

    Article  CAS  Google Scholar 

  109. H. Yumerefendi, F. Tarendeau, P. J. Mas and D. J. Hart, J Struct Biol., 172, 66 (2010).

    Article  CAS  Google Scholar 

  110. Y. An, H. Yumerefendi, P. J. Mas, A. Chesneau and D. J. Hart, J. Struct. Biol., 175, 189 (2011).

    Article  CAS  Google Scholar 

  111. B. Nölting, Methods in modern biophysics: Ch. 5 — protein infrared spectroscopy, Springer Berlin Heidelberg, 91 (2010).

  112. R. L. Wiseman and J.W. Kelly, Mol. Cell., 36, 730 (2009).

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Chemical Engineering, University of Virginia, Charlottesville, Virginia, 22904, USA

    Simpson Gregoire, Jacob Irwin & Inchan Kwon

  2. Institutes on Aging, University of Virginia, Charlottesville, Virginia, 22904, USA

    Inchan Kwon

Authors
  1. Simpson Gregoire
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jacob Irwin
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Inchan Kwon
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Inchan Kwon.

Additional information

Inchan Kwon is Assistant Professor of the Chemical Engineering at the University of Virginia. Professor Kwon earned his BS and MS in Chemical Engineering at Seoul National University, South Korea (BS and MS). He received his PhD in Chemical Engineering from California Institute of Technology (Caltech) and completed his postdoctoral training in the Bioengineering at the University of California at Berkley. Professor Kwon focuses on the engineering proteins using nonnatural amino acid incorporation and the modulation of protein aggregation associated with neurodegenerative diseases including Alzheimer’s disease and Lou Gehrig’s disease. Professor Kwon has been recognized in numerous ways including Constantin G. Economou Memorial Award at Caltech, KSEA Young Investigator Grant, and James M. Lee Young Investigator Award.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gregoire, S., Irwin, J. & Kwon, I. Techniques for monitoring protein misfolding and aggregation in vitro and in living cells. Korean J. Chem. Eng. 29, 693–702 (2012). https://doi.org/10.1007/s11814-012-0060-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11814-012-0060-x

Key words

Search

Navigation