Skip to main content
SpringerLink
Log in

Inhibitor analysis of protein phosphorylation/dephosphorylation in maize mitochondria in relation to redox conditions

  • Research Papers
  • Published:
Russian Journal of Plant Physiology Aims and scope Submit manuscript

Abstract

The role of protein phosphorylation/ dephosphorylation in the redox regulation of mitochondrial functioning was investigated. Incubation of isolated mitochondria of maize (Zea mays L.) in the presence of γ-32P-ATP revealed phosphorylation of polypeptides with mol wt of 66, 60, 55, 48/50 doublet, 45, 29, 22, and 19 kD. The presence in the incubation medium of oxidized glutathione significantly reduced the level of protein phosphorylation. The addition of reduced glutathione diminished phosphorylation of proteins with mol wt of 60 and 48/50 kD and slightly increased phosphorylation of proteins with mol wt of 66, 55, and 45 kD. The reducing agent, sodium dithionite decreased phosphorylation of proteins with mol wt of 60, 45, 29, 22, and 19 kD but increased phosphorylation of 55 kD protein. The inhibitors of protein kinases and protein phosphatases significantly modified the effects of redox agents. For example, simultaneous action of an oxidant K3[Fe(CN)6] and NaF enhanced phosphorylation level compared to separate treatments with these agents. The combined application of sodium dithionite and NaF elevated phosphorylation level of 55 kD protein. Phosphoprotein with mol wt of about 66 kD was identified immunochemically as a heat shock protein (HSP 60). The results indicate the presence in mitochondria of redox-sensitive protein kinases and protein phosphatases. Differential changes in the pattern of mitochondrial phosphoproteins under the action of various redox agents suggest that phosphorylation is probably involved in the transduction of redox signal in plant mitochondria.

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

Abbreviations

GSH:

reduced glutathione

GSSG:

oxidized glutathione

HSP:

heat shock proteins

MPT:

mitochondrial permeability transition

PTP:

protein tyrosine phosphatases

References

  1. Schwartz, R.M. and Dayhoff, M.O., Origins of Prokaryotes, Eukaryotes, Mitochondria, and Chloroplasts, Science, 1978, vol. 199, pp. 395–403.

    Article  PubMed  CAS  Google Scholar 

  2. Allen, J.F., Alexciev, K., and Hakansson, G., Regulation by Redox Signalling, Curr. Biol., 1995, vol. 5, pp. 869–872.

    Article  PubMed  CAS  Google Scholar 

  3. Allen, R.G. and Tresini, M., Oxidative Stress and Gene Regulation, Free Radic. Biol., 2000, vol. 28, pp. 463–499.

    Article  CAS  Google Scholar 

  4. Karin, M., Signal Transduction from the Cell Surface to the Nucleus through the Phosphorylation of Transcription Factors, Curr. Opin. Cell Biol., 1994, vol. 6, pp. 415–424.

    Article  PubMed  CAS  Google Scholar 

  5. Litchfield, D.W., Protein Kinase CK2: Structure, Regulation and Role in Cellular Decisions of Life and Death, Biochem. J., 2003, vol. 369, pp. 1–15.

    Article  PubMed  CAS  Google Scholar 

  6. Whitmarsh, A.J. and Davis, R.J., Regulation of Transcription Factor Function by Phosphorylation, Cell Mol. Life Sci., 2000, vol. 57, pp. 1172–1183.

    Article  PubMed  CAS  Google Scholar 

  7. El-Kady, A. and Klenova, E., Regulation of the Transcription Factor, CTCF, by Phosphorylation with Protein Kinase CK2, FEBS Lett., 2005, vol. 579, pp. 1424–1434.

    Article  PubMed  CAS  Google Scholar 

  8. Prieto-Martin, A., Montoya, J., and Martinez-Asorin, F., Phosphorylation of Rat Mitochondrial Transcription Termination Factor (mTEFA) Is Required for Transcription Termination but Not for Binding to DNA, Nucleic Acids Res., 2004, vol. 32, pp. 2059–2068.

    Article  PubMed  CAS  Google Scholar 

  9. El’skaya, A.V., Starodub, N.F., Potapov, A.P., Kovalenko, M.I., Ovcharenko, G.V., Obolenskaya, M.Yu., and Ivanov, L.L., Regulyatsiya biosinteza belka u eukariot (Regulation of Protein Biosynthesis in Eukaryotes), Kiev: Naukova Dumka, 1990.

    Google Scholar 

  10. Arrigo, A.-P., Gene Expression and the Thiol Redox State, Free Radic. Biol., 1999, vol. 27, pp. 936–944.

    Article  CAS  Google Scholar 

  11. Ohno, S., Akita, Y., Konno, Y., Imajoh, S., and Suzuki, K., A Novel Phorbol Ester Receptor/Protein Kinase, nPKC, Distantly Related to the Protein Kinase C Family, Cell, 1988, vol. 53, pp. 731–741.

    Article  PubMed  CAS  Google Scholar 

  12. Monteiro, H.P. and Stern, A., Redox Regulation of Tyrosine Phosphorylation-Dependent Signal Transduction Pathway, Free Radic. Biol. Med., 1996, vol. 21, pp. 323–333.

    Article  PubMed  CAS  Google Scholar 

  13. Bykova, N.V., Egsgaard, H., and Moller, I.M., Identification of 14 New Phosphoproteins Involved in Important Plant Mitochondrial Processes, FEBS Lett., 2003, vol. 540, pp. 141–146.

    Article  PubMed  CAS  Google Scholar 

  14. Subota, I.Yu., Arziev, A.Sh., and Konstantinov, Yu.M., The Involvement of Protein Phosphorylation/Dephosphorylation in the Redox Control of Translation in Cereal Mitochondria, Russ. J. Plant Physiol., 2004, vol. 51, pp. 784–789.

    Article  CAS  Google Scholar 

  15. Konstantinov, Yu.M., Lutsenko, G.N., and Podsosonny, V.A., Inhibition of Adenine Nucleotide Translocation in Maize Seedling Mitochondria by Anionic Detergents, Physiol. Plant., 1988, vol. 72, pp. 403–406.

    Article  CAS  Google Scholar 

  16. Struglics, A., Fredlund, K.M., Konstantinov, Yu.M., Allen, J.F., and Moller, I.M., Protein Phosphorylation/Dephosphorylation in the Inner Membrane of Potato Tuber Mitochondria, FEBS Lett., 2000, vol. 475, pp. 213–217.

    Article  PubMed  CAS  Google Scholar 

  17. Laemmli, U.K., Cleavage of Structural Proteins during the Assembly of Head of Bacteriophage T4, Nature, 1970, vol. 277, pp. 174–182.

    Google Scholar 

  18. Timmons, T.M. and Dunbar, B.S., Protein Blotting and Immunodetection, Methods Enzymol., 1990, vol. 182, pp. 679–701.

    Article  PubMed  CAS  Google Scholar 

  19. Combet, C., Blanchet, C., Geourjon, C., and Deleage, G., NPS: Network Protein Sequence Analysis, Trends Biochem. Sci., 2000, vol. 25, pp. 147–150.

    Article  PubMed  CAS  Google Scholar 

  20. Konstantinov, Yu.M., Subota, I.Yu., and Arziev, A.Sh., Effects of Glutathione Redox-System on Translation Activity in Zea mays, Dokl. Akad. Nauk, 1999, vol. 365, pp. 126–128.

    CAS  Google Scholar 

  21. Baena-Gonzales, E., Baginsky, S., Mulo, P., Summer, H., Aro, E.-M., and Link, G., Chloroplast Transcription at Different Light Intensities: Glutathione-Mediated Phosphorylation of the Major RNA Polymerase Involved in Redox-Regulation Organellar Gene Expression, Plant Physiol., 2001, vol. 127, pp. 1044–1052.

    Article  Google Scholar 

  22. Elizarov, S.M., Michurina, T.A., and Danilenko, V.N., Proteinase Activity of Serine/Threonine in Cell-Depleted Extracts of Streptomyces lividans, Antibiot. Khimioter., 1998, no. 2, pp. 3–8.

  23. Huang, J., Wei, Y., Kim, Y., Octerberg, L., and Matthews, H.R., Purification of Protein Kinase from the Yeast Saccharomyces cerevisiae, J. Biol. Chem., 1991, vol. 266, pp. 9023–9031.

    PubMed  CAS  Google Scholar 

  24. Prasad, T.K. and Hallberg, R.L., cDNA Clones Encoding Arabidopsis thaliana and Zea mays Mitochondrial Chaperonin HSP60 and Gene Expression during Seed Germination and Heat Shock, Plant Mol. Biol., 1992, vol. 18, pp. 873–885.

    Article  PubMed  CAS  Google Scholar 

  25. Lund, A.A., Blum, P.H., Bhattramakki, D., and Elthon, T.E., Heat-Stress Response of Maize Mitochondria, Plant Physiol., 1998, vol. 116, pp. 1097–1110.

    Article  PubMed  CAS  Google Scholar 

  26. Hochholdinger, F., Guo, L., and Schnable, P.S., Cytoplasmic Regulation of the Accumulation of Nuclear-Encoded Proteins in the Mitochondrial Proteome of Maize, Plant J., 2004, vol. 37, pp. 199–208.

    PubMed  CAS  Google Scholar 

  27. Khan, I.V., Wallin, R., Gupta, R.S., and Kammer, G.M., Protein Kinase A-Catalyzed Phosphorylation of Heat Shock Protein 60 Chaperone Regulates Its Attachment to Histone 2B in the T Lymphocyte Plasma Membrane, Proc. Natl. Acad. Sci. USA, 1998, vol. 95, pp. 10425–10430.

    Article  PubMed  CAS  Google Scholar 

  28. Cvoro, A., Dundjerski, J., Trajkovic, D., and Matic, G., The Level and Phosphorylation of HSP70 in the Rat Liver Cytosol after Adrenalectomy and Hyperthermia, Cell Biol. Int., 1999, vol. 23, pp. 313–320.

    Article  PubMed  CAS  Google Scholar 

  29. Azarashvili, T.S., Tyynela, J., Odinokova, I.V., Grigorjev, P.A., Baumann, M., Evtodienko, Y.V., and Saris, N.-E., Phosphorylation of a Peptide Related to Subunite C of the F0F1-ATPase/ATP Synthase and Relationship to Permeability Transition Pore Opening in Mitochondria, J. Bioenerg. Biomembr., 2002, vol. 34, pp. 279–284.

    Article  PubMed  CAS  Google Scholar 

  30. Arpagaus, S., Rawyler, A.J., and Braendle, R.A., Occurrence and Characteristics of the Mitochondrial Permeability Transition in Plants, J. Biol. Chem., 2002, vol. 277, pp. 1780–1787.

    Article  PubMed  CAS  Google Scholar 

  31. Petrussa, E., Casolo, V., Braidot, E., Chiandussi, E., Macri, F., and Vianello, A., Cyclosporin A Induces the Opening of a Potassium-Selective Channel in Higher Plant Mitochondria, J. Bioenerg. Biomembr., 2001, vol. 33, pp. 107–117.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Siberian Institute of Plant Physiology and Biochemistry, Siberian Division, Russian Academy of Sciences, a/ya 317, ul. Lermontova 132, Irkutsk, 664033, Russia

    I. Yu. Subota, A. Sh. Arziev, L. P. Senzhenko, V. I. Tarasenko & Yu. M. Konstantinov

Authors
  1. I. Yu. Subota
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. Sh. Arziev
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. L. P. Senzhenko
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. V. I. Tarasenko
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yu. M. Konstantinov
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Original Russian Text © I.Yu. Subota, A.Sh. Arziev, L.P. Senzhenko, V.I. Tarasenko, Yu.M. Konstantinov, 2007, published in Fiziologiya Rastenii, 2007, Vol. 54, No. 3, pp. 389–396.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Subota, I.Y., Arziev, A.S., Senzhenko, L.P. et al. Inhibitor analysis of protein phosphorylation/dephosphorylation in maize mitochondria in relation to redox conditions. Russ J Plant Physiol 54, 343–349 (2007). https://doi.org/10.1134/S1021443707030089

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S1021443707030089

Key words

Search

Navigation