Extended Data Fig. 5: Chromatography assay of CaMKII sensors.
From: Dendritic, delayed, stochastic CaMKII activation in behavioural time scale plasticity
To estimate the approximate oligomeric state of CaMKII sensors, we performed FSEC on EGFP-CaMKIIα and various Camui proteins expressed in HEK293 cells by transient transfection. The cell lysates expressing these proteins were directly injected to a Superose-6 column, and EGFP fluorescence was detected using 475/507 nm excitation/emission wavelengths. Green Camuiα (1dV-Camui), Green Camuiβ (in which CaMKIIβ subunit is used instead of CaMKIIα), and 2dV-Camui are compared with EGFP-CaMKIIα. 2dV-Camui showed a similar retention time to EGFP-CaMKIIα and 1dV-Camui (peak at ~27 min; arrow 1), suggesting the formation of a dodecamer. However, Green Camuiβ showed a high fraction of lower oligomeric species represented by a peak at ~35 min (arrow 2).
