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Background

αCaMKII plays central and essential roles in long-term potentiation (LTP), learning and memory. αCaMKII is activated via binding with Ca²⁺/CaM in response to elevated Ca²⁺ concentration. Furthermore, prolonged increase in Ca²⁺ concentration leads to the auto-phosphorylation of αCaMKII at T286, maintaining the activation of αCaMKII even after Ca²⁺/CaM dissociation. Importantly, the active form of αCaMKII is thought to exhibit conformational change. In order to elucidate the relationships between the interaction of αCaMKII with CaM and the conformational change of αCaMKII, we generated molecular probes (YFP-αCaMKII with CFP-CaM and YFP-αCaMKII-CFP) and performed time-lapse imaging of the interaction with CaM and the conformational change, respectively, in living cells using FRET.

Results

The interaction of YFP-αCaMKII with CFP-CaM and the conformational change of YFP-αCaMKII-CFP were induced simultaneously in response to increased concentrations of Ca²⁺. Consistent with previous predictions, high levels of Ca²⁺ signaling maintained the conformational change of YFP-αCaMKII-CFP at the time when CFP-CaM was released from YFP-αCaMKII. These observations indicated the transfer of αCaMKII conformational change from CaM-dependence to CaM-independence. Furthermore, analyses using αCaMKII mutants showed that phosphorylation at T286 and T305/306 played positive and negative roles, respectively, during in vivo interaction with CaM and further suggested that CaM-dependent and CaM-independent conformational changed forms displays similar but distinct structures.

Conclusions

Importantly, these structual differences between CaM-dependent and -independent forms of αCaMKII may exhibit differential functions for αCaMKII, such as interactions with other molecules required for LTP and memory. Our molecular probes could thus be used to identify therapeutic targets for cognitive disorders that are associated with the misregulation of αCaMKII."xsd:string
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http://purl.uniprot.org/citations/23958294http://purl.uniprot.org/core/author"Kato K."xsd:string
http://purl.uniprot.org/citations/23958294http://purl.uniprot.org/core/author"Iwamoto T."xsd:string
http://purl.uniprot.org/citations/23958294http://purl.uniprot.org/core/author"Kida S."xsd:string
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http://purl.uniprot.org/citations/23958294http://purl.uniprot.org/core/title"Interactions between alphaCaMKII and calmodulin in living cells: conformational changes arising from CaM-dependent and -independent relationships."xsd:string
http://purl.uniprot.org/citations/23958294http://purl.uniprot.org/core/volume"6"xsd:string
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