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Protein kinase activity in Cucumis sativus cotyledons: Effect of calcium and light
The effect of calcium and light on the protein phosphorylation and protein kinase activity of Cucumis sativus cotyledons was studied in order to investigate phytochrome-dependence and to understand phytochrome signal transduction better. Light signals received by phytochromes in plants may be transd...
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Published in: | Phytochemistry (Oxford) 2007-02, Vol.68 (4), p.438-445 |
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Main Authors: | , , , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
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Summary: | The effect of calcium and light on the protein phosphorylation and protein kinase activity of
Cucumis sativus cotyledons was studied in order to investigate phytochrome-dependence and to understand phytochrome signal transduction better.
Light signals received by phytochromes in plants may be transduced through protein phosphorylation. Ca
2+ as second messenger was involved in phytochrome-mediated cellular events. Our experiments with
Cucumis sativus cotyledons, treated with red (R) and far-red (FR) light, showed a stimulatory effect on
in vitro protein phosphorylation of histone, added as exogenous substrate to the cotyledon extracts, and also modified the phosphorylation of endogenous polypeptides. The effect of light treatments was mimicked by the addition of Ca
2+ to the phosphorylation buffer, indicating phytochrome- and Ca
2+-dependence on activity of some protein kinases (PKs). In-gel kinase assays were performed to characterize the PKs involved at the cotyledon stage of cucumber plants. Three proteins of about 75, 57 and 47
kDa with PK activity were detected between
M
r markers of 94 and 45
kDa. All three were able to phosphorylate histone and undergo autophosphorylation. However, only the 75 and 57
kDa proteins autophosphorylated and phosphorylated the substrate in a Ca
2+-dependent manner, and were inhibited when calmodulin (CaM) antagonists were added to the incubation buffer. Western-blot analysis with polyclonal antibodies directed against calcium-dependent protein kinase of rice (OsCDPK11) or
Arabidopsis (AtCPK2) recognised 57 and 75
kDa polypeptides, respectively. These results indicate the presence in cucumber cotyledons of at least two proteins (ca. 75 and 57
kDa) with activity of PKs that could be calcium-dependent protein kinases (CDPKs). Both CDPKs could be modulated by phytochromes throughout FR-HIR and VLFR responses. |
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ISSN: | 0031-9422 1873-3700 |
DOI: | 10.1016/j.phytochem.2006.11.001 |