Molecular cloning of a full-length cDNA for ECBP21 from

Guohong, Mao

doi:10.1360/02tb9247

Cited by 2 publications

(3 citation statements)

References 19 publications

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“…Tang et al (1996) have determined that purified native ECBP21 protein binds to CaM in the Ca 2+ -dependent manner. In addition, the recombinant ECBP21 also binds to CaM in the presence of calcium (Mao et al 2002). To map the CaM binding region of ECBP21, two truncated proteins were prepared.…”

Section: Discussionmentioning

confidence: 99%

“…The full-length cDNA coding for ECBP21 was isolated according to the procedure of Mao et al (2002). Purified ECBP21 from A. dahurica suspension-cultured cells was subjected to SDS-PAGE and electroblotted onto PVDF membrane.…”

Section: Plant Materialsmentioning

confidence: 99%

“…In our previous studies, a full-length cDNA coding for ECBP21 was isolated and identified (Mao et al 2002). In this work, GenBank database research was performed for comparison and alignment of the amino acid sequence with known protein sequences.…”

Section: Introductionmentioning

confidence: 99%

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Characterization of a cDNA coding for an extracellular calmodulin-binding protein from suspension-cultured cells of Angelica dahurica

Guohong

Hou

Ding

et al. 2005

Planta

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In order to characterize a specific extracellular 21-kDa calmodulin-binding protein (named: ECBP21) from Angelica dahurica L. suspension-cultured cells, the cDNA coding for the protein has been cloned. Here, Southern blot analysis shows that there are at least two copies of ECBP21 gene in Angelica genome. Using truncated versions of ECBP21 and synthetic peptide in CaM binding assays, we mapped the calmodulin-binding domain to a 16-amino acid stretch (residues 200-215) at the C-terminal region. The ECBP21 was localized in the cell wall area by the immunogold electron microscopy and by GFP labeling method. These results define ECBP21 as a kind of an extracellular calmodulin-binding protein (CaMBP). Furthermore, using Northern blot analysis, we examined the expression dynamics of ecbp21 during the incubation of Angelica suspension-cultured cells and the treatments with some growth regulators. The above studies further provide the molecular evidence for the existence of the gene coding for extracellular CaMBPs and imply a possible role for ECBP21.

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Section: Discussionmentioning

confidence: 99%

Section: Plant Materialsmentioning

confidence: 99%

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Characterization of a cDNA coding for an extracellular calmodulin-binding protein from suspension-cultured cells of Angelica dahurica

Guohong

Hou

Ding

et al. 2005

Planta

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Oxidative defence reactions in sunflower roots induced by methyl-jasmonate and methyl-salicylate and their relation with calcium signalling

2009

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Ca(2+) plays a critical role as second messenger in the signal-response coupling of plant defence responses, and methyl-jasmonate and methyl-salicylate are important components of signal transduction cascades activating plant defences. When intact axenic non-induced seedling roots of sunflower were treated with different Ca(2+) concentrations up to 1 mM, there was no significant increase in O(2)(*-) generation or DMAB-MBTH peroxidase (extracellular, ECPOX) activities in the apoplast, probably because these roots had enough Ca(2+) in their exo- and endocellular reservoirs. Both activities were strongly inhibited by the RBOH-NADPH oxidase inhibitor DPI and by the Ca(2+) surrogate antagonist La(3+), but the voltage-dependent Ca(2+) channel blocker verapamil was only inhibitory at concentrations higher than those active on animal L-type Ca(2+) channels. Concentrations >5 mM EGTA (chelating Ca(2+) in the apoplast) and Li(+) (inhibiting PI cycle dependent endogenous Ca(2+) fluxes) also inhibited both activities. W7, inhibitor of binding of Ca-CaM to its target protein, enhanced both activities, but the inactive analogue W5 showed a similar effect. Our data suggest that Ca(2+) from exocellular and, to a lesser extent, from endocellular stores is involved in oxidative activities, and that RBOH-NADPH oxidase is the main system supporting them. Ca(2+) activation of the PM cytosolic side of RBOH-NADPH oxidase is probably the key to Ca(2+) involvement in these processes. Roots induced by MeJA or MeSA showed significant enhancement of both oxidative activities, as corresponding to the oxidative burst evoked by the two phytohormones in the root apoplast. But while ECPOX activity showed a response to the effectors similar to that described above for non-induced roots, O(2)(*-) generation activity in the apoplast of induced roots was insensitive to EGTA, verapamil and Li(+), the inhibitors of exogenous and endogenous Ca(2+) fluxes; only DPI and La(3+) were inhibitory. As exogenously added 0.1 mM Ca(2+) also increased O (2) (.-) generation, we propose that, in these roots, activation of RBOH-NADPH oxidase by Ca(2+) could be regulated by Ca(2+) sensors in the apoplast.

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Molecular cloning of a full-length cDNA for ECBP21 from

Cited by 2 publications

References 19 publications

Characterization of a cDNA coding for an extracellular calmodulin-binding protein from suspension-cultured cells of Angelica dahurica

Characterization of a cDNA coding for an extracellular calmodulin-binding protein from suspension-cultured cells of Angelica dahurica

Oxidative defence reactions in sunflower roots induced by methyl-jasmonate and methyl-salicylate and their relation with calcium signalling

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