Advertisement

Method for Protein Subfractionation of Cardiovascular Tissues Before DIGE Analysis

  • Athanasios Didangelos
  • Xiaoke Yin
  • Manuel MayrEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 854)

Abstract

Difference gel electrophoresis (DIGE) (Electrophoresis 18, 2071–2077, 1997, 1) is widely used in cardiovascular research. However, the dynamic range limitations stemming from contaminating plasma proteins and highly abundant extracellular matrix components can make cardiovascular tissues difficult to analyze. Here we describe a novel methodology for biochemical subfractionation of cardiovascular tissues before DIGE analysis.

Key words

Cardiovascular Heart Proteomics Vascular Vessel 

Notes

Acknowledgements

M.M. is a senior fellow of the British Heart Foundation.

References

  1. 1.
    Unlu, M., Morgan, M. E., and Minden, J. S. (1997) Difference gel electrophoresis: a single gel method for detecting changes in protein extracts, Electrophoresis 18, 2071–2077.PubMedCrossRefGoogle Scholar
  2. 2.
    Arrell, D. K., Neverova, I., and Van Eyk, J. E. (2001) Cardiovascular proteomics: evolution and potential, Circ Res 88, 763–773.PubMedCrossRefGoogle Scholar
  3. 3.
    Didangelos, A., Simper, D., Monaco, C., and Mayr, M. (2009) Proteomics of acute coronary syndromes, Curr Atheroscler Rep 11, 188–195.PubMedCrossRefGoogle Scholar
  4. 4.
    Mayr, M., Liem, D., Zhang, J., Li, X., Avliyakulov, N. K., Yang, J. I., Young, G., Vondriska, T. M., Ladroue, C., Madhu, B., Griffiths, J. R., Gomes, A., Xu, Q., and Ping, P. (2009) Proteomic and metabolomic analysis of cardioprotection: Interplay between protein kinase C epsilon and delta in regulating glucose metabolism of murine hearts, J Mol Cell Cardiol 46, 268–277.PubMedCrossRefGoogle Scholar
  5. 5.
    Mayr, M., Madhu, B., and Xu, Q. (2007) Proteomics and metabolomics combined in cardiovascular research, Trends Cardiovasc Med 17, 43–48.PubMedCrossRefGoogle Scholar
  6. 6.
    Mayr, M., Zhang, J., Greene, A. S., Gutterman, D., Perloff, J., and Ping, P. (2006) Proteomics-based development of biomarkers in cardiovascular disease: mechanistic, clinical, and therapeutic insights, Mol Cell Proteomics 5, 1853–1864.PubMedCrossRefGoogle Scholar
  7. 7.
    McGregor, E., and Dunn, M. J. (2006) Proteomics of the heart: unraveling disease, Circ Res 98, 309–321.PubMedCrossRefGoogle Scholar
  8. 8.
    Yin, X., Cuello, F., Mayr, U., Hao, Z., Hornshaw, M., Ehler, E., Avkiran, M., and Mayr, M. (2010) Proteomics analysis of the cardiac myofilament subproteome reveals dynamic alterations in phosphatase subunit distribution, Mol Cell Proteomics 9, 497–509.PubMedCrossRefGoogle Scholar
  9. 9.
    Didangelos, A., Yin, X., Mandal, K., Baumert, M., Jahangiri, M., and Mayr, M. (2010) Proteomics characterization of extracellular space components in the human aorta, Mol Cell Proteomics 9, 2048–2062.PubMedCrossRefGoogle Scholar
  10. 10.
    Mason, R. M., and Mayes, R. W. (1973) Extraction of cartilage protein-polysaccharides with inorganic salt solutions, Biochem J 131, 535–540.PubMedGoogle Scholar
  11. 11.
    Dahl, S. L., Koh, J., Prabhakar, V., and Niklason, L. E. (2003) Decellularized native and engineered arterial scaffolds for transplantation, Cell Transplant 12, 659–666.PubMedGoogle Scholar
  12. 12.
    Korossis, S. A., Wilcox, H. E., Watterson, K. G., Kearney, J. N., Ingham, E., and Fisher, J. (2005) In-vitro assessment of the functional performance of the decellularized intact porcine aortic root, J Heart Valve Dis 14, 408–421; discussion 422.Google Scholar
  13. 13.
    Gilbert, T. W., Sellaro, T. L., and Badylak, S. F. (2006) Decellularization of tissues and organs, Biomaterials 27, 3675–3683.PubMedGoogle Scholar
  14. 14.
    Sajdera, S. W., and Hascall, V. C. (1969) Proteinpolysaccharide complex from bovine nasal cartilage. A comparison of low and high shear extraction procedures, J Biol Chem 244, 77–87.Google Scholar
  15. 15.
    Bannister, D. W., and Burns, A. B. (1972) Pepsin treatment of avian skin collagen. Effects on solubility, subunit composition and aggregation properties, Biochem J 129, 677–681.Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Athanasios Didangelos
    • 1
  • Xiaoke Yin
    • 1
  • Manuel Mayr
    • 1
    Email author
  1. 1.Cardiovascular DivisionKing’s College LondonLondonUK

Personalised recommendations