• Molecules and Cells
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• v.24 no.2
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• pp.276-282
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• 2007

Protein phosphorylation is one of the major mechanisms by which eukaryotic cells transduce extracellular signals into intracellular responses. Calcium/calmodulin ($Ca^{2+}/CaM$)-dependent protein phosphorylation has been implicated in various cellular processes, yet little is known about $Ca^{2+}/CaM$-dependent protein kinases (CaMKs) in plants. From an Arabidopsis expression library screen using a horseradish peroxidase-conjugated soybean calmodulin isoform (SCaM-1) as a probe, we isolated a full-length cDNA clone that encodes AtCK (Arabidopsis thaliana calcium/calmodulin-dependent protein kinase). The predicted structure of AtCK contains a serine/threonine protein kinase catalytic domain followed by a putative calmodulin-binding domain and a putative $Ca^{2+}$-binding domain. Recombinant AtCK was expressed in E. coli and bound to calmodulin in a $Ca^{2+}$-dependent manner. The ability of CaM to bind to AtCK was confirmed by gel mobility shift and competition assays. AtCK exhibited its highest levels of autophosphorylation in the presence of 3 mM $Mn^{2+}$. The phosphorylation of myelin basic protein (MBP) by AtCK was enhanced when AtCK was under the control of calcium-bound CaM, as previously observed for other $Ca^{2+}/CaM$-dependent protein kinases. In contrast to maize and tobacco CCaMKs (calcium and $Ca^{2+}/CaM$-dependent protein kinase), increasing the concentration of calmodulin to more than $3{\mu}M$ suppressed the phosphorylation activity of AtCK. Taken together our results indicate that AtCK is a novel Arabidopsis $Ca^{2+}/CaM$-dependent protein kinase which is presumably involved in CaM-mediated signaling.

• Journal of Plant Biotechnology
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• v.30 no.4
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• pp.399-403
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• 2003

In order to understand the protection effects of antioxidant enzymes against oxidative stress caused by various environmental stresses, transgenic tobacco (Nicotiana tabacum cv, Xanthi) plants expressing both copper/zinc superoxide dismutase (CuZnSOD) and ascorbate peroxidase (APX) in chloroplasts (referred to as CA plants) were subjected to highlight (1,100$\mu$mol m$^{-2}$ sec$^{-1}$) and chilling at 4$^{\circ}C$. The protection effects of CA plants using leaf discs were compared with those of transgenic plants expressing either CuZnSOD or APX in chloroplasts (SOD plants or APX plants, respectively) and non-transgenic (NT) plants. CA plants showed about 15％ protection in the photosynthetic efficiency (Fv/Fm) of photosystem II relative to NT plants 1 hr after treatment of both highlight and chilling, whereas they showed about 23％ protection in the redox state of P700 in photosystem I at 3 hr after treatment. SOD plants or APX plants showed an intermediate protection effect between CA plants and NT plants. These results demonstrated that the coexpression of CuZnSOD and APX in chloroplasts importantly involves in the protection effects against oxidative stress caused by various environmental stresses.