TRACKING OF SINGLE FLUORESCENTLY TAGGED CAMKII

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Ca2+/calmodulin-dependent protein kinase II (CaMKII) is a multimeric complex composed predominantly of two subunits, alpha and beta. This holoenzyme, with a radius of gyration of ~20nm, consists of twelve subunits and plays an important role in synaptic plasticity. During synaptic activation calcium enters dendritic spines where it loads onto calmodulin (CaM) that subsequently activates CaMKII. The activated kinase moves to the postsynaptic density, where it is thought to phosphorylate AMPA receptors, thus potentiating synaptic transmission. These qualitative observations have yet to be backed-up through quantitative analysis of kinase movement. We propose to monitor the traceability of fluorescently labeled CaMKII, with twelve fluorescent proteins (eGFP) attached, would allow us to quantify the effect of diffusion during this activation pathway.