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Preparation of a peptide template for the SRM method by means of in vitro phosphorylation

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Biochemistry (Moscow) Supplement Series B: Biomedical Chemistry Aims and scope Submit manuscript

Abstract

Phosphorylation is one of the most common and the most important posttranslational modifications (PTM) of proteins. The study of protein phosphoforms is complicated by their low abundance in analyzed biological samples compared with the abundance of corresponding unmodified proteins. The method of selected reactions monitoring (SRM) allows to perform very sensitive and selective PTM analysis with high specificity and sensitivity. Using myelin basic protein (MBP) as a model we have developed a method for phosphoprotein detection by SRM. The method is based on obtaining phosphoproteins in a reconstituted kinase system and following usage of tryptic fragments of the phosphorylated protein as a template for the development of the SRM method. The developed method has been successfully applied for detection of phosphopeptides of myelin basic protein in human brain glioma samples.

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References

  1. Khoury, G.A., Baliban, R.C., and Floudas, C.A., Scientific Reports, 2011, vol. 1, p. 90. doi: 10.1038/srep00090

    Article  CAS  Google Scholar 

  2. Cohen, P., Trends Biochem. Sci., 2000, vol. 25, pp. 596–601. doi: 10.1016/S0968-0004(00)01712-6

    Article  CAS  Google Scholar 

  3. Hunter, T., Cell, 2000, vol. 100, pp. 113–127. doi: 10.1016/S0092-8674(00)81688-8

    Article  CAS  Google Scholar 

  4. Wrighton, K.H. and Feng, X.-H., Cellular Signalling, 2008, vol. 20, pp. 1579–1591. doi: 10.1016/jcellsig.2008.02.003

    Article  CAS  Google Scholar 

  5. Olsen, J.V., Blagoev, B., Gnad, F., Macek, B., Kumar, C., Mortensen, P., and Mann, M., Cell, 2006, vol. 127, pp. 635–648. doi: 10.1016/jcell.2006.09.026

    Article  CAS  Google Scholar 

  6. Mann, M., Ong, S.-E., Grønborg, M., Steen, H., Jensen, O.N., and Pandey, A., Trends Biotech., 2002, vol. 20, pp. 261–268. doi: 10.1016/S01677799(02)01944-3

    Article  CAS  Google Scholar 

  7. Gu, B. and Zhu, W.-G., Int. J. Biol. Sci., 2012, vol. 8, pp. 672–684. doi: 10.7150/ijbs.4283

    Article  Google Scholar 

  8. Wrighton, K.H., Lin, X., and Feng, X.-H., Cell Res., 2009, vol. 19, pp. 8–20. doi: 10.1038/cr.2008.327

    Article  CAS  Google Scholar 

  9. Katsogiannou, M., Andrieu, C., and Rocchi, P., Frontiers in Genetics, 2014, vol. 5, p. 346. doi: 10.3389/fgene.2014.00346

    Article  Google Scholar 

  10. Kelleher, N.L., Analyt. Chem., 2004, vol. 76, pp. 196 A–203 A. doi: 10.1021/ac0415657

    Article  Google Scholar 

  11. Dunn, J.D., Reid, G.E., Bruening, M.L., Mass Spectrometry Rev., 2010, vol. 29, pp. 29–54. doi: 10.1002/mas.20219

    CAS  Google Scholar 

  12. Jin, L.L., Tong, J., Prakash, A., Peterman, S.M., StGermain, J.R., Taylor, P., and Moran, M.F., J. Proteome Res., 2010, vol. 9, pp. 2752–2761. doi: 10.1021/pr100024a

    Article  CAS  Google Scholar 

  13. Wolf-Yadlin, A., Hautaniemi, S., Lauffenburger, D.A., and White, F.M., Proc. Natl. Acad. Sci. USA, 2007, vol. 104, pp. 5860–5865. doi: 10.1073/pnas.0608638104

    Article  CAS  Google Scholar 

  14. Anderson, L. and Hunter, C.L., Mol. Cell Proteomics: MCP, 2006, vol. 5, pp. 573–588. doi: 10.1074/mcp.M500331-MCP200

    Article  CAS  Google Scholar 

  15. Sherrod, S.D., Myers, M.V., Li, M., Myers, J.S., Carpenter, K.L., Maclean, B., and Ham, A.-J.L., J. Proteome Res., 2012, vol. 11, pp. 3467–3479. doi: 10.1021/pr201240a

    Article  CAS  Google Scholar 

  16. Peterson, A.C., Russell, J.D., Bailey, D.J., Westphall, M.S., and Coon, J.J., Mol. Cell Proteomics: MCP, 2012, vol. 11, pp. 1475–1488. doi: 10.1074/mcp.O112.020131

    Article  Google Scholar 

  17. Gallien, S., Duriez, E., Crone, C., Kellmann, M., Moehring, T., and Domon, B., Mol. Cell Proteomics: MCP, 2012, vol. 11, pp. 1709–1723. doi: 10.1074/mcp.O112.019802

    Article  Google Scholar 

  18. http://worldwidepromegacom/resources/protocols/product-information-sheets/n/erk2-kinase-enzymesystem-protocol/

  19. https://wwwbiochemmpgde/226356/FASP

  20. Walker, J.M., The Protein Protocols Handbook, 2002, pp. 11–14. doi: 10.1385/1-59259-169-8:11

    Book  Google Scholar 

  21. Vassall, K.A., Bessonov, K., De Avila, M., Polverini, E., and Harauz, G., PLoS One, 2013, vol. 8, e68175. doi: 10.1371/journalpone.0068175

    Article  Google Scholar 

  22. Liu, Y., Hüttenhain, R., Collins, B., and Aebersold, R., Expert Rev. Mol. Diagn., 2013, vol. 13, pp. 811–825. doi: 10.1586/14737159.2013.845089

    Article  CAS  Google Scholar 

  23. Kim, J.K., Mastronardi, F.G., Wood, D.D., Lubman, D.M., Zand, R., and Moscarello, M.A., Mol. Cell Proteomics: MCP, 2003, vol. 2, pp. 453–462. doi: 10.1074/mcp.M200050-MCP200

    CAS  Google Scholar 

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Authors and Affiliations

  1. Institute of Biomedical Chemistry (IBMC), ul. Pogodinskaya 10, Moscow, 119121, Russia

    M. G. Zavialova, V. G. Zgoda, O. N. Kharybin & E. N. Nikolayev

  2. Institute for Energy Problems of Chemical Physics of the Russian Academy of Sciences, Moscow, Russia

    E. N. Nikolayev

Authors
  1. M. G. Zavialova
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  2. V. G. Zgoda
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  3. O. N. Kharybin
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  4. E. N. Nikolayev
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Corresponding author

Correspondence to V. G. Zgoda.

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Original Russian Text © M.G. Zavialova, V.G. Zgoda, O.N. Kharybin, E.N. Nikolayev, 2015, published in Biomeditsinskaya Khimiya.

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Zavialova, M.G., Zgoda, V.G., Kharybin, O.N. et al. Preparation of a peptide template for the SRM method by means of in vitro phosphorylation. Biochem. Moscow Suppl. Ser. B 9, 343–350 (2015). https://doi.org/10.1134/S1990750815040095

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  • DOI: https://doi.org/10.1134/S1990750815040095

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