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Literature summary for 2.7.11.17 extracted from

  • Miller, J.B.; Pratap, A.; Miyahara, A.; Zhou, L.; Bornemann, S.; Morris, R.J.; Oldroyd, G.E.
    Calcium/calmodulin-dependent protein kinase is negatively and positively regulated by calcium, providing a mechanism for decoding calcium responses during symbiosis signaling (2013), Plant Cell, 25, 5053-5066.
    View publication on PubMedView publication on EuropePMC

Activating Compound

Activating Compound Comment Organism Structure
additional information the enzyme is activated by calmdulin binding at higher Ca2+ concentrations, autoactivation of CCaMK by mutation of Thr271 Medicago truncatula

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of MBP-fusion wild-type enzyme and of His-tagged mutant enzymes in Escherichia coli strain BL21 (DE3), expression of recombinant GST-tagged wild-type enzyme in Escherichia coli strain Rosetta (DE3), recombinant expression of the kinase domain of CCaMK alone gives rise to spontaneous nodulation 8 weeks after root transformation, but in the absence of rhizobia Medicago truncatula

Protein Variants

Protein Variants Comment Organism
E319A site-directed mutagenesis, Thr271 phosphorylation is reduced in the mutant compared to the wild-type Medicago truncatula
L324A site-directed mutagenesis, Thr271 phosphorylation is abolished in the L324A mutant compared to the wild-type Medicago truncatula
L333A site-directed mutagenesis, Thr271 phosphorylation is enhanced in the L324A mutant compared to the wild-type, resulting in a constitutively active enzyme Medicago truncatula
R323A site-directed mutagenesis, the mutant is unable to autoactivate Medicago truncatula
S322A site-directed mutagenesis, the mutant is able to autoactivate Medicago truncatula
S343A site-directed mutagenesis, Thr271 phosphorylation is reduced in the mutant compared to the wild-type Medicago truncatula
T271A site-directed mutagenesis, the mutant is able to autoactivate Medicago truncatula
T271D site-directed mutagenesis, the mutant is able to autoactivate Medicago truncatula
T271I site-directed mutagenesis, the mutant is able to autoactivate Medicago truncatula
T271X autoactivation of CCaMK by mutation of Thr271 Medicago truncatula

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ required for binding of cofactor calmodulin, differential calcium binding affinities of the EF-hand domains of the enzyme. Ca2+ binding via the EF-hand domains promotes Thr271 phosphorylation and enzyme deactivation, a deactivating conformation of CCaMK is stabilized by Thr271 phosphorylation. The binding of Ca2+ negatively regulates CCaMK by enhancing Thr271 phosphorylation Medicago truncatula
Mg2+ required Medicago truncatula

Organism

Organism UniProt Comment Textmining
Medicago truncatula Q6RET7 cv. Jemalong A17
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Posttranslational Modification

Posttranslational Modification Comment Organism
phosphoprotein enzyme phosphorylation at residue Thr271, CCaMK autophosphorylation modeling, overview Medicago truncatula

Purification (Commentary)

Purification (Comment) Organism
recombinant tagged wild-type enzymes from Escherichia coli strains by affinity chromatography, recombinant His-tagged mutant enzymes from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography and gel filtration Medicago truncatula

Subunits

Subunits Comment Organism
More modelling of two stable states for CCaMK: an inactive form that predominates at basal Ca2+ concentrations and involves a hydrogen bond network between the CaM binding and kinase domains and an active form that predominates during Ca2+ oscillations associated with CaM binding Medicago truncatula

Synonyms

Synonyms Comment Organism
calcium/calmodulin-dependent protein kinase
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Medicago truncatula
CCaMK
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Medicago truncatula
ccamk-1
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Medicago truncatula
DMI-3
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Medicago truncatula
dmi3-1
-
Medicago truncatula
TRV25
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Medicago truncatula

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Medicago truncatula

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Medicago truncatula

Cofactor

Cofactor Comment Organism Structure
ATP
-
Medicago truncatula
Calmodulin required, calcium-dependent CaM binding overrides the effects of autophosphorylation and activates the protein Medicago truncatula

General Information

General Information Comment Organism
malfunction autoactivation of CCaMK by mutation of Thr271. The complete absence of the EF-hand domains leads to unregulated Thr271 phosphorylation and deactivation of the protein Medicago truncatula
additional information modelling of two stable states for CCaMK: an inactive form that predominates at basal Ca2+ concentrations and involves a hydrogen bond network between the CaM binding and kinase domains and an active form that predominates during Ca2+ oscillations associated with CaM binding. The hydrogen bond between phosphorylated Thr271 and Ser322 is the predominant stabilizing bond at the inactive fold in Medicago truncatula CCaMK Medicago truncatula
physiological function the differential calcium binding affinities of the EF-hand domains compared with those of calmodulin suggest that enzyme CCaMK is maintained in the inactive state at basal calcium concentrations and is activated via calmodulin binding during calcium oscillations. Recombinant expression of the kinase domain of CCaMK alone gives rise to spontaneous nodulation 8 weeks after root transformation, but in the absence of rhizobia. The binding of Ca2+ negatively regulates CCaMK by enhancing Thr271 phosphorylation, while binding of calmodulin activates it, the enzyme remains inactive at basal Ca2+ concentrations and becomes activated during Ca2+ spiking. Enzyme regulation modelling, overview Medicago truncatula