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Effect of epibrassinolide on tyrosine phosphorylation of the calvin cycle enzymes

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Abstract

Two-dimensional electrophoresis was used to separate proteins from crude extracts of pea (Pisum sativum L.) leaves, and thus isolated proteins were subjected to Western blot analysis with monoclonal antibodies against PY20 phosphotyrosine polypeptides. This analysis revealed 44 polypeptides phosphorylated on tyrosine residues. Phosphorylation of some of these proteins was changed under the action of epibrassinolide. Some of these polypeptides were identified by means of MALDI-TOF MS analysis. The results indicate that eight of these proteins belong to the Calvin cycle enzymes, namely, the isoforms of Rubisco large and small subunits, fructose-1,6-phosphate aldolases 1 and 2, and the precursor of α-subunit of Rubisco-binding protein. The observed changes in phosphorylation of these proteins may partly explain the effects of brassinosteroids on photosynthesis. The tyrosine phosphorylation sites were identified in silico for the fragments of polypeptides examined.

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Abbreviations

2D electrophoresis:

two-dimensional electrophoresis

CHAPS:

3-[(3-cholamidepropyl)-dimethylammonio]-1-propanesulfonate

DTT:

dithiothreitol

EPB:

24-epibrassinolide

IEF:

isoelectric focusing

MALDI-TOF MS:

matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

PMSF:

phenylmethylsulfonyl fluoride

PVDF membranes:

polyvinyldifluoride membranes

TBST:

Tris buffered saline supplemented with Tween-20

TEMED:

tetramethylethylenediamine

TFA:

trifluoroacetic acid

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Authors and Affiliations

  1. Kazan Institute of Biochemistry and Biophysics, Kazan Research Center, Russian Academy of Sciences, Kazan, a/ya 30, Tatarstan, 420111, Russia

    E. O. Fedina, F. G. Karimova & I. A. Tarchevsky

  2. Orekhovich Institute of Biomedical Chemistry, Russian Academy of Medical Sciences, Moscow, Russia

    I. Yu. Toropygin

  3. Institute of Bioorganic Chemistry, National Academy of Sciences of Belarus, Minsk, Belarus

    V. A. Khripach

Authors
  1. E. O. Fedina
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  2. F. G. Karimova
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  3. I. A. Tarchevsky
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  4. I. Yu. Toropygin
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  5. V. A. Khripach
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Correspondence to E. O. Fedina.

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Original Russian Text © E.O. Fedina, F.G. Karimova, I.A. Tarchevsky, I.Yu. Toropygin, V.A. Khripach, 2008, published in Fiziologiya Rastenii, 2008, Vol. 55, No. 2, pp. 210–218.

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Fedina, E.O., Karimova, F.G., Tarchevsky, I.A. et al. Effect of epibrassinolide on tyrosine phosphorylation of the calvin cycle enzymes. Russ J Plant Physiol 55, 193–200 (2008). https://doi.org/10.1134/S1021443708020052

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  • DOI: https://doi.org/10.1134/S1021443708020052

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