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Applied Biochemistry and Biotechnology

, Volume 163, Issue 1, pp 64–70 | Cite as

Use of Highly Purified and Mixed Antibodies for Simultaneous Detection of Multiple Protein Species Released from Mitochondria upon Induction of the Permeability Transition

  • Takenori Yamamoto
  • Mizuki Ohashi
  • Sho Mizutani
  • Yuuya Yoshimura
  • Eriko Obana
  • Hiroshi Terada
  • Yasuo ShinoharaEmail author
Article
  • 93 Downloads

Abstract

Concomitant with the induction of the mitochondrial permeability transition (PT), cytochrome c is released from mitochondria into the cytosol where it triggers subsequent steps of cellular apoptosis. Thus, inducers of the mitochondrial PT would become “seed compounds” of regulators of apoptosis. However, when we examine the actions of certain chemicals on the release of mitochondrial cytochrome c, the behaviors of not only cytochrome c but also multiple mitochondrial protein species must be carefully examined because the mitochondrial PT and release of proteins from mitochondria occur in diverse manners. In the present study, we examined whether it is possible to measure the behaviors of multiple protein species in a single experiment using purified and mixed antibodies. The results obtained clearly indicate that this procedure would be applicable for high-throughput screening of regulators of apoptosis. Further requirements necessary for the establishment of a useful screening system for apoptosis regulators are discussed.

Keywords

Mitochondria Permeability transition Protein release Cytochrome c Western blotting 

References

  1. 1.
    Crompton, M. (1999). Biochemical Journal, 341, 233–249.CrossRefGoogle Scholar
  2. 2.
    Kroemer, G., & Reed, J. C. (2000). Nature Medicine, 6, 513–519.CrossRefGoogle Scholar
  3. 3.
    Bernardi, P., Petronilli, V., Di Lisa, F., & Forte, M. (2001). Trends in Biochemical Sciences, 26, 112–117.CrossRefGoogle Scholar
  4. 4.
    Gunter, T. E., & Pfeiffer, D. R. (1990). American Journal of Physiology: Cell Physiology, 258, C755–C786.Google Scholar
  5. 5.
    Zoratti, M., & Szabò, I. (1995). Biochimica et Biophysica Acta, 1241, 139–176.Google Scholar
  6. 6.
    Bernardi, P., Colonna, R., Costantini, P., Eriksson, O., Fontaine, E., Ichas, F., et al. (1998). Biofactors, 8, 273–281.CrossRefGoogle Scholar
  7. 7.
    Shinohara, Y., Bandou, S., Kora, S., Kitamura, S., Inazumi, S., & Terada, H. (1998). FEBS Letters, 428, 89–92.CrossRefGoogle Scholar
  8. 8.
    Almofti, M. R., Ichikawa, T., Yamashita, K., Terada, H., & Shinohara, Y. (2003). Journal of Biochemistry, 134, 43–49.CrossRefGoogle Scholar
  9. 9.
    Yamamoto, T., Tachikawa, A., Terauchi, S., Yamashita, K., Kataoka, M., Terada, H., et al. (2004). European Journal of Biochemistry, 271, 3573–3579.CrossRefGoogle Scholar
  10. 10.
    Yamamoto, T., Terauchi, S., Tachikawa, A., Yamashita, K., Kataoka, M., Terada, H., et al. (2005). Journal of Bioenergetics and Biomembranes, 37, 299–306.CrossRefGoogle Scholar
  11. 11.
    Yamamoto, T., Yoshimura, Y., Yamada, A., Gouda, S., Yamashita, K., Yamazaki, N., et al. (2008). Journal of Bioenergetics and Biomembranes, 40, 619–623.CrossRefGoogle Scholar
  12. 12.
    Shinohara, Y., Almofti, M. R., Yamamoto, T., Ishida, T., Kita, F., Kanzaki, H., et al. (2002). European Journal of Biochemistry, 269, 5224–5230.CrossRefGoogle Scholar
  13. 13.
    Yamada, A., Yamamoto, T., Yamazaki, N., Yamashita, K., Kataoka, M., Nagata, T., et al. (2009). Molecular & Cellular Proteomics, 8, 1265–1277.CrossRefGoogle Scholar
  14. 14.
    Terauchi, S., Yamamoto, T., Yamashita, K., Kataoka, M., Terada, H., & Shinohara, Y. (2005). Mitochondrion, 5, 248–254.CrossRefGoogle Scholar
  15. 15.
    Tzul, F. O., Kurchan, E., Roder, H., & Bowler, B. E. (2009). Biochemistry, 48, 481–491.CrossRefGoogle Scholar
  16. 16.
    Scarlett, J. L., & Murphy, M. P. (1997). FEBS Letters, 418, 282–286.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Takenori Yamamoto
    • 1
  • Mizuki Ohashi
    • 1
    • 2
  • Sho Mizutani
    • 1
    • 2
  • Yuuya Yoshimura
    • 1
    • 2
  • Eriko Obana
    • 1
    • 2
  • Hiroshi Terada
    • 3
  • Yasuo Shinohara
    • 1
    • 2
    Email author
  1. 1.Institute for Genome ResearchUniversity of TokushimaTokushimaJapan
  2. 2.Faculty of Pharmaceutical SciencesUniversity of TokushimaTokushimaJapan
  3. 3.Faculty of Pharmaceutical SciencesTokyo University of ScienceNodaJapan

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